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Alexa Fluor® 700 anti-mouse I-A/I-E Antibody Alexa Fluor® 700 anti-mouse I-A/I-E Antibody

Pricing & Availability
Clone
M5/114.15.2 (See other available formats)
Regulatory Status
RUO
Other Names
MHC class II
Isotype
Rat IgG2b, κ
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Product Citations
publications
1_M5slash114_Alx700_020608
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I-E (clone M5/114.15.2)Alexa Fluor® 700 (filled histogram) or rat IgG2b, κ Alexa Fluor® 700 isotype control (open histogram).
  • 1_M5slash114_Alx700_020608
    C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I-E (clone M5/114.15.2)Alexa Fluor® 700 (filled histogram) or rat IgG2b, κ Alexa Fluor® 700 isotype control (open histogram).
  • 34_Mouse_Spleen_F480_MHCII
    Confocal image of C57BL/6 mouse spleen sample acquired using the IBEX method of highly multiplexed antibody-based imaging: MHCII (blue) in Cycle 2 and F4/80 (magenta) in Cycle 2. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 39_Mouse_Thymus_CD68_MHCII
    Confocal image of C57BL/6 mouse thymus sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD68 (cyan) in Cycle 3 and MHCII (red) in Cycle 3. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 45_Mouse_Lung_EpCAM_MHCII_CD169
    Confocal image of C57BL/6 mouse lung sample acquired using the IBEX method of highly multiplexed antibody-based imaging: EpCAM (blue) in Cycle 2, MHCII (magenta) in Cycle 2, and CD169 (green) in Cycle 4. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 51_Mouse_Gut_EpCAM_CD31_MHCII
    Confocal image of C57BL/6 mouse small intestine sample acquired using the IBEX method of highly multiplexed antibody-based imaging: EpCAM (blue) in Cycle 1, CD31 (magenta) in Cycle 2, and MHCII (green) in Cycle 3. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 55_Mouse_Liver_Ecadherin_MHCII
    Confocal image of C57BL/6 mouse liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: E-Cadherin (blue) in Cycle 1 and MHCII (magenta) in Cycle 4. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Compare all formats See Alexa Fluor® 700 spectral data
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107621 25 µg DKK559
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107622 100 µg DKK1278
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Description

These class II molecules are expressed on antigen presenting cells (including B cells) and a subset of T cells from H-2b,d,q,r bearing mice and are involved in antigen presentation to T cells expressing CD3/TCR and CD4 proteins.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Activated C57BL/6 mouse spleen cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 700 under optimal conditions.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested

SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. The suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. It is highly recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 700 has a maximum emission of 719 nm when it is excited at 633 nm / 635 nm. Prior to using Alexa Fluor® 700 conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

View full statement regarding label licenses
Excitation Laser
Red Laser (633 nm)
Application Notes

The M5/114.15.2 antibody reacts with a polymorphic determinant shared by the I-Ab, I-Ad, I-Aq, I-Ed, and I-Ek MHC class II alloantigens from mice carrying H-2p,r,q,b,d,u haplotypes. Clone M5/114.15.2 however does not react wtih I-Af, I-Ak, or I-As MHC class II alloantigens.1

Additional reported applications (for the relevant formats) include: immunoprecipitation1, immunohistochemistry of frozen sections2,3,6, in vitro and in vivo blocking of antigen presentation or ligand binding4-7, and spatial biology (IBEX)17,18. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. Nos. 107655 & 107656).

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References
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  2. Viville S, et al. 1993. Cell 72:635. (IHC)
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  4. Shi Y, et al. 1998. J. Exp. Med. 187:367. (Block)
  5. Yamashita I, et al. 1993. Int. Immunol. 5:1139.
  6. Guo M, et al. 1995. Zygote 3:65. (IHC)
  7. Kim A, et al. 2004. Exp. Mol. Med. 36:428. (Block)
  8. Luckashenak NA, et al. 2006. J. Immunol. 177:5177.
  9. Venanzi ES, et al. 2007. J. Immunol. 179:5693.
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  17. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  18. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
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  127. Hindy G, et al. 2022. J Clin Invest. 132: . PubMed
  128. Mansouri S, et al. 2022. J Immunol. 209:2114. PubMed
  129. Tatsumi N, et al. 2022. STAR Protoc. 3:101845. PubMed
  130. Cousin N, et al. 2022. Cells. 12: . PubMed
  131. Sieow JL, et al. 2023. Int J Mol Sci. 24: . PubMed
  132. Zhao K, et al. 2023. Front Immunol. 14:1101769. PubMed
  133. Jameson VJ, et al. 2022. Cytometry A. 101:922. PubMed
  134. Bohrer AC, et al. 2022. Cell Rep. 40:111144. PubMed
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  136. Bonifacio JPPL, et al. 2022. J Virol. 96:e0087122. PubMed
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  139. Solier S, et al. 2023. Nature. 617:386. PubMed
  140. Huang T, et al. 2023. Cell Death Dis. 14:354. PubMed
  141. Mathur R, et al. 2019. Mucosal Immunol. 12:612. PubMed
  142. Hou X, et al. 2020. Cell Reports. 28(1):172-189.e7.. PubMed
  143. Komuczki J, et al. 2019. Immunity. 50:1289. PubMed
  144. Kwong Chung CK, et al. 2017. Immunology. 150:221. PubMed
  145. Huang W, et al. 2015. J Allergy Clin Immunol. . PubMed
  146. Liu Y, et al. 2021. Cell Metabolism. 33(6):1221-1233.e11. PubMed
  147. Zhang R, et al. 2019. Cell Rep. 28:2647. PubMed
  148. Mancini M, et al. 2019. J Immunol. 202:1479. PubMed
  149. Wiesner DL, et al. 2020. Cell Host Microbe. 614:27. PubMed
  150. Hewitson JP, et al. 2020. J Immunol. 204:2949. PubMed
  151. Baptista AP et al. 2019. Immunity. 50(5):1188-1201 . PubMed
  152. Yilmaz B, et al. 2021. Cell Host Microbe. 29(4):650-663.e9. PubMed
  153. Uzhachenko RV, et al. 2021. Cell Reports. 35(1):108944. PubMed
  154. Dong L, et al. 2021. Cancer Cell. . PubMed
  155. Tordesillas L, et al. 2018. Nat Commun. 9:5238. PubMed
  156. Tyagi AM et al. 2018. Immunity. 49(6):1116-1131 . PubMed
  157. Pack AD, et al. 2021. Cell Reports. 36:109586. PubMed
  158. Hearnden R, et al. 2021. STAR Protocols. 2(2):100422. PubMed
  159. Earnest JT, et al. 2021. Cell Reports. 35(1):108962. PubMed
  160. Chikina AS, et al. 2020. Cell. 183:411. PubMed
  161. Zhang F, et al. 2019. Nat Commun. 10:3974. PubMed
  162. Chan YH, et al. 2019. EMBO Mol Med. 11:e10092. PubMed
  163. Dupont C, et al. 2015. J Immunol. 195: 4369 - 4377. PubMed
  164. Webster HC, et al. 2020. J Immunol Methods. 112702:477. PubMed
  165. Go DM, et al. 2021. Cell Mol Gastroenterol Hepatol. 12:715. PubMed
  166. Hering L, et al. 2021. Int J Mol Sci. 22:. PubMed
  167. Cousin N, et al. 2021. Cancer Res. 81:4133. PubMed
  168. Salei N, et al. 2020. J Am Soc Nephrol. 31:257. PubMed
  169. Uher O, et al. 2021. Cancers (Basel). 13:. PubMed
  170. Wiesner D, et al. 2015. PLoS Pathog. 11:1004701. PubMed
  171. Ku MW, et al. 2020. Cell Host Microbe. 29(2):236-249.e6. PubMed
  172. Trompette A, et al. 2022. Mucosal Immunol. :. PubMed
  173. Mansouri S, et al. 2021. J Immunol. 206:2233. PubMed
  174. Benechet AP, et al. 2019. Nature. 574:200. PubMed
  175. Mrdjen D et al. 2018. Immunity. 48(2):380-395 . PubMed
  176. Korangath P, et al. 2020. Bio Protoc. 10:e3822. PubMed
  177. Ruhland MK, et al. 2020. Cancer Cell. 37(6):786-799.e5. PubMed
  178. Ortiz-Bonilla CJ, et al. 2022. Int J Mol Sci. 23:. PubMed
  179. Leylek R, et al. 2019. Cell Rep. 29:3736. PubMed
  180. Sutherland TE, et al. 2021. Frontiers in Immunology. 12:715209. PubMed
  181. Brandi P, et al. 2022. Cell Rep. 38:110184. PubMed
  182. , et al. 2021. Eur J Immunol. 51:2708. PubMed
  183. Cohen SB et al. 2018. Cell host & microbe. 24(3):439-446 . PubMed
  184. Steuerman Y et al. 2018. Cell systems. 6(6):679-691 . PubMed
  185. Witkowski MT, et al. 2020. Cancer Cell. 37:867. PubMed
  186. Tacconi C, et al. 2021. Cell Reports. 35(2):108993. PubMed
  187. Dieterich LC, et al. 2020. Angiogenesis. 1.24375. PubMed
  188. Dupont C, et al. 2014. PLoS Pathog. 10:1004047. PubMed
  189. Ndeupen S, et al. 2021. iScience. 24:103479. PubMed
  190. Rhys HI, et al. 2018. EBioMedicine. 29:60. PubMed
  191. Macdougall CE et al. 2018. Cell metabolism. 27(3):588-601 . PubMed
  192. Martínez‐López M et al. 2019. Immunity. 50(2):446-461 . PubMed
  193. Casulli J, et al. 2019. Nat Commun. 10:2121. PubMed
  194. Chen YG, et al. 2020. Molecular Cell. 76(1):96-109. PubMed
  195. Liu S, et al. 2020. Cell Host & Microbe. 26(6):779-794.e8.. PubMed
  196. Sharma A, et al. 2020. Cell. 183(2):377-394.e21. PubMed
  197. Deerhake ME, et al. 2021. Immunity. 54(3):484-498.e8. PubMed
  198. De Simone G, et al. 2021. Immunity. :. PubMed
  199. Kyburz A, et al. 2019. J Allergy Clin Immunol. 143:1496. PubMed
  200. Janela B, et al. 2019. Immunity. 50:1069. PubMed
  201. Blriot C, et al. 2021. Immunity. :. PubMed
  202. Liu X, et al. 2020. Cell Host Microbe. 28(5):683-698.e6. PubMed
  203. Zhang X, et al. 2021. Mol Cancer Res. 19:1076. PubMed
  204. Gomez S, et al. 2022. J Immunother Cancer. 10:. PubMed
  205. Toshiro Hirai et al. 2019. Immunity. 50(5):1249-1261 . PubMed
  206. Camara A, et al. 2019. Immunity. 50:1467. PubMed
  207. Koelwyn GJ, et al. 2020. Nat Med. 1452:26. PubMed
  208. Kim DK, et al. 2022. Nat Commun. 13:6292. PubMed
  209. Quiding-Järbrink M, et al. 2010. PLoS One. 5:e15018. PubMed
  210. Xueyang Yu et al. 2017. Immunity. 47(5):903-912 . PubMed
  211. Cunha LD et al. 2018. Cell. 175(2):429-441 . PubMed
  212. Zeis P, et al. 2020. Immunity. 53:775. PubMed
  213. Desai P, et al. 2021. Cell. 184(5):1214-1231.e16. PubMed
  214. Mansouri S, et al. 2020. Mucosal Immunol. 0.954861111. PubMed
  215. Teo T, et al. 2015. J Virol. 89: 7955-7969. PubMed
  216. Draijer C, et al. 2018. Sci Rep. 8:5105. PubMed
  217. Andersen TK, et al. 2019. NPJ Vaccines. 4:9. PubMed
  218. Yousef H, et al. 2019. Nat Med. 25:988. PubMed
  219. Utz SG, et al. 2020. Cell. 181(3):557-573. PubMed
  220. Varikuti S, et al. 2020. Br J Cancer. 122:1005. PubMed
  221. Winkler ES, et al. 2020. Cell. 182(4):901-918.e18. PubMed
  222. Leary N, et al. 2022. J Extracell Vesicles. 11:e12197. PubMed
  223. Salazar V, et al. 2019. Cell Rep. 26:1585. PubMed
  224. Montalban-Arques A, et al. 2021. Cell Host Microbe. :. PubMed
  225. Meunier I, et al. 2015. PLoS One. 10: 0138055. PubMed
  226. Mujal AM, et al. 2022. Cancer Immunol Res. 10:403. PubMed
  227. Hoyer FF, et al. 2020. Immunity. 51(5):899-914.e7.. PubMed
  228. Huang WC, et al. 2020. Adv Mater. 32:e2005637. PubMed
  229. Hering L, et al. 2020. Front Immunol. 1.747222222. PubMed
  230. Santopaolo M, et al. 2021. Front Immunol. 609406:12. PubMed
  231. Ceglia V, et al. 2021. J Immunol. 207:2060. PubMed
  232. Schmit T, et al. 2022. Cell Rep. 38:110456. PubMed
  233. Mogilenko DA et al. 2019. Cell. 177(5):1201-1216 . PubMed
  234. Braza MS et al. 2018. Immunity. 49(5):819-828 . PubMed
  235. Bonavita E, et al. 2020. Immunity. 1215:53. PubMed
  236. Ubags ND, et al. 2021. J Allergy Clin Immunol. 1049:147. PubMed
  237. Droho S, et al. 2020. J Vis Exp. . PubMed
  238. Srivastava S, et al. 2020. Cancer Cell. 39(2):193-208.e10. PubMed
  239. Spiljar M, et al. 2021. Cell Metab. 33:2231. PubMed
  240. Mansouri S, et al. 2021. J Immunol. 206:2233. PubMed
  241. Rivera CA, et al. 2022. Immunity. 55:129. PubMed
  242. Wu L, et al. 2020. Cancer Immunol Res. 710:8. PubMed
  243. Lee S, et al. 2021. Autophagy. 18:1062. PubMed
  244. Syed I et al. 2018. Cell metabolism. 27(2):419-427 . PubMed
  245. Aurélien Trompette et al. 2018. Immunity. 48(5):992-1005 . PubMed
  246. C Khouili S, et al. 2020. Cell Rep. 33:108468. PubMed
RRID
AB_493726 (BioLegend Cat. No. 107621)
AB_493727 (BioLegend Cat. No. 107622)

Antigen Details

Structure
MHC class II
Distribution

B cell and activated T cells, APCs of the H-2b,d,q,r bearing mice

Function
Antigen presentation
Ligand/Receptor
CD3/TCR, CD4
Cell Type
Antigen-presenting cells, B cells, Dendritic cells, T cells, Tregs
Biology Area
Immunology, Innate Immunity
Molecular Family
MHC Antigens
Antigen References

1. Watts C. 1997. Ann. Rev. Immunol. 15:821.
2. Pamer E, et al. 1998. Ann. Rev. Immunol. 16:323.

Gene ID
14961 View all products for this Gene ID 14969 View all products for this Gene ID
UniProt
View information about I-A/I-E on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All I-A/I-E Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-mouse I-A/I-E (Maxpar® Ready) M5/114.15.2 FC,CyTOF®
Brilliant Violet 605™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 650™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 711™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 785™ anti-mouse I-A/I-E M5/114.15.2 FC
PE/Dazzle™ 594 anti-mouse I-A/I-E M5/114.15.2 FC
Alexa Fluor® 594 anti-mouse I-A/I-E M5/114.15.2 IHC-F,3D IHC
APC/Fire™ 750 anti-mouse I-A/I-E M5/114.15.2 FC
TotalSeq™-A0117 anti-mouse I-A/I-E M5/114.15.2 PG
Ultra-LEAF™ Purified anti-mouse I-A/I-E M5/114.15.2 FC,IHC-F,IP,Block
Spark Blue™ 550 anti-mouse I-A/I-E M5/114.15.2 FC
TotalSeq™-B0117 anti-mouse I-A/I-E M5/114.15.2 PG
PE/Fire™ 640 anti-mouse I-A/I-E M5/114.15.2 FC
Biotin anti-mouse I-A/I-E M5/114.15.2 FC
FITC anti-mouse I-A/I-E M5/114.15.2 FC
PE anti-mouse I-A/I-E M5/114.15.2 FC
Purified anti-mouse I-A/I-E M5/114.15.2 FC,IHC-F,IP,Block
TotalSeq™-C0117 anti-mouse I-A/I-E M5/114.15.2 PG
PE/Cyanine5 anti-mouse I-A/I-E M5/114.15.2 FC
APC anti-mouse I-A/I-E M5/114.15.2 FC
Alexa Fluor® 488 anti-mouse I-A/I-E M5/114.15.2 FC,3D IHC
Alexa Fluor® 647 anti-mouse I-A/I-E M5/114.15.2 FC
Pacific Blue™ anti-mouse I-A/I-E M5/114.15.2 FC
Alexa Fluor® 700 anti-mouse I-A/I-E M5/114.15.2 FC,SB
PerCP/Cyanine5.5 anti-mouse I-A/I-E M5/114.15.2 FC
PerCP anti-mouse I-A/I-E M5/114.15.2 FC
APC/Cyanine7 anti-mouse I-A/I-E M5/114.15.2 FC
PE/Cyanine7 anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 421™ anti-mouse I-A/I-E M5/114.15.2 FC,IHC-F,SB
Brilliant Violet 510™ anti-mouse I-A/I-E M5/114.15.2 FC
Spark YG™ 581 anti-mouse I-A/I-E M5/114.15.2 FC
PE/Fire™ 810 anti-mouse I-A/I-E M5/114.15.2 FC
Spark UV™ 387 anti-mouse I-A/I-E M5/114.15.2 FC
Spark Violet™ 538 anti-mouse I-A/I-E M5/114.15.2 FC
PerCP/Fire™ 806 anti-mouse I-A/I-E M5/114.15.2 FC
Spark Red™ 718 anti-mouse I-A/I-E M5/114.15.2 FC
APC/Fire™ 810 anti-mouse I-A/I-E M5/114.15.2 FC
Spark PLUS UV395™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 750™ anti-mouse I-A/I-E M5/114.15.2 FC
Spark YG™ 593 anti-mouse I-A/I-E (Flexi-Fluor™) Antibody M5/114.15.2 FC
Spark NIR™ 685 anti-mouse I-A/I-E (Flexi-Fluor™) Antibody M5/114.15.2 FC
StarBright UltraViolet 795 anti-mouse I-A/I-E M5/114.15.2 FC
Spark PLUS B574™ anti-mouse I-A/I-E Antibody M5/114.15.2 FC
Go To Top Version: 6    Revision Date: 04/21/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

Pricing & Availability
Clone
M5/114.15.2 (See other available formats)
Regulatory Status
RUO
Other Names
MHC class II
Isotype
Rat IgG2b, κ
Ave. Rating
Submit a Review
Product Citations
publications
1_M5slash114_Alx700_020608
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I-E (clone M5/114.15.2)Alexa Fluor® 700 (filled histogram) or rat IgG2b, κ Alexa Fluor® 700 isotype control (open histogram).
  • 1_M5slash114_Alx700_020608
    C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I-E (clone M5/114.15.2)Alexa Fluor® 700 (filled histogram) or rat IgG2b, κ Alexa Fluor® 700 isotype control (open histogram).
  • 34_Mouse_Spleen_F480_MHCII
    Confocal image of C57BL/6 mouse spleen sample acquired using the IBEX method of highly multiplexed antibody-based imaging: MHCII (blue) in Cycle 2 and F4/80 (magenta) in Cycle 2. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 39_Mouse_Thymus_CD68_MHCII
    Confocal image of C57BL/6 mouse thymus sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD68 (cyan) in Cycle 3 and MHCII (red) in Cycle 3. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 45_Mouse_Lung_EpCAM_MHCII_CD169
    Confocal image of C57BL/6 mouse lung sample acquired using the IBEX method of highly multiplexed antibody-based imaging: EpCAM (blue) in Cycle 2, MHCII (magenta) in Cycle 2, and CD169 (green) in Cycle 4. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 51_Mouse_Gut_EpCAM_CD31_MHCII
    Confocal image of C57BL/6 mouse small intestine sample acquired using the IBEX method of highly multiplexed antibody-based imaging: EpCAM (blue) in Cycle 1, CD31 (magenta) in Cycle 2, and MHCII (green) in Cycle 3. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 55_Mouse_Liver_Ecadherin_MHCII
    Confocal image of C57BL/6 mouse liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: E-Cadherin (blue) in Cycle 1 and MHCII (magenta) in Cycle 4. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Compare all formats See Alexa Fluor® 700 spectral data
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107621 25 µg DKK559
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107622 100 µg DKK1278
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Description

These class II molecules are expressed on antigen presenting cells (including B cells) and a subset of T cells from H-2b,d,q,r bearing mice and are involved in antigen presentation to T cells expressing CD3/TCR and CD4 proteins.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Activated C57BL/6 mouse spleen cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 700 under optimal conditions.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested

SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. The suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. It is highly recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 700 has a maximum emission of 719 nm when it is excited at 633 nm / 635 nm. Prior to using Alexa Fluor® 700 conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

View full statement regarding label licenses
Excitation Laser
Red Laser (633 nm)
Application Notes

The M5/114.15.2 antibody reacts with a polymorphic determinant shared by the I-Ab, I-Ad, I-Aq, I-Ed, and I-Ek MHC class II alloantigens from mice carrying H-2p,r,q,b,d,u haplotypes. Clone M5/114.15.2 however does not react wtih I-Af, I-Ak, or I-As MHC class II alloantigens.1

Additional reported applications (for the relevant formats) include: immunoprecipitation1, immunohistochemistry of frozen sections2,3,6, in vitro and in vivo blocking of antigen presentation or ligand binding4-7, and spatial biology (IBEX)17,18. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. Nos. 107655 & 107656).

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References
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  17. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  18. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
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RRID
AB_493726 (BioLegend Cat. No. 107621)
AB_493727 (BioLegend Cat. No. 107622)

Antigen Details

Structure
MHC class II
Distribution

B cell and activated T cells, APCs of the H-2b,d,q,r bearing mice

Function
Antigen presentation
Ligand/Receptor
CD3/TCR, CD4
Cell Type
Antigen-presenting cells, B cells, Dendritic cells, T cells, Tregs
Biology Area
Immunology, Innate Immunity
Molecular Family
MHC Antigens
Antigen References

1. Watts C. 1997. Ann. Rev. Immunol. 15:821.
2. Pamer E, et al. 1998. Ann. Rev. Immunol. 16:323.

Gene ID
14961 View all products for this Gene ID 14969 View all products for this Gene ID
UniProt
View information about I-A/I-E on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All I-A/I-E Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-mouse I-A/I-E (Maxpar® Ready) M5/114.15.2 FC,CyTOF®
Brilliant Violet 605™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 650™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 711™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 785™ anti-mouse I-A/I-E M5/114.15.2 FC
PE/Dazzle™ 594 anti-mouse I-A/I-E M5/114.15.2 FC
Alexa Fluor® 594 anti-mouse I-A/I-E M5/114.15.2 IHC-F,3D IHC
APC/Fire™ 750 anti-mouse I-A/I-E M5/114.15.2 FC
TotalSeq™-A0117 anti-mouse I-A/I-E M5/114.15.2 PG
Ultra-LEAF™ Purified anti-mouse I-A/I-E M5/114.15.2 FC,IHC-F,IP,Block
Spark Blue™ 550 anti-mouse I-A/I-E M5/114.15.2 FC
TotalSeq™-B0117 anti-mouse I-A/I-E M5/114.15.2 PG
PE/Fire™ 640 anti-mouse I-A/I-E M5/114.15.2 FC
Biotin anti-mouse I-A/I-E M5/114.15.2 FC
FITC anti-mouse I-A/I-E M5/114.15.2 FC
PE anti-mouse I-A/I-E M5/114.15.2 FC
Purified anti-mouse I-A/I-E M5/114.15.2 FC,IHC-F,IP,Block
TotalSeq™-C0117 anti-mouse I-A/I-E M5/114.15.2 PG
PE/Cyanine5 anti-mouse I-A/I-E M5/114.15.2 FC
APC anti-mouse I-A/I-E M5/114.15.2 FC
Alexa Fluor® 488 anti-mouse I-A/I-E M5/114.15.2 FC,3D IHC
Alexa Fluor® 647 anti-mouse I-A/I-E M5/114.15.2 FC
Pacific Blue™ anti-mouse I-A/I-E M5/114.15.2 FC
Alexa Fluor® 700 anti-mouse I-A/I-E M5/114.15.2 FC,SB
PerCP/Cyanine5.5 anti-mouse I-A/I-E M5/114.15.2 FC
PerCP anti-mouse I-A/I-E M5/114.15.2 FC
APC/Cyanine7 anti-mouse I-A/I-E M5/114.15.2 FC
PE/Cyanine7 anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 421™ anti-mouse I-A/I-E M5/114.15.2 FC,IHC-F,SB
Brilliant Violet 510™ anti-mouse I-A/I-E M5/114.15.2 FC
Spark YG™ 581 anti-mouse I-A/I-E M5/114.15.2 FC
PE/Fire™ 810 anti-mouse I-A/I-E M5/114.15.2 FC
Spark UV™ 387 anti-mouse I-A/I-E M5/114.15.2 FC
Spark Violet™ 538 anti-mouse I-A/I-E M5/114.15.2 FC
PerCP/Fire™ 806 anti-mouse I-A/I-E M5/114.15.2 FC
Spark Red™ 718 anti-mouse I-A/I-E M5/114.15.2 FC
APC/Fire™ 810 anti-mouse I-A/I-E M5/114.15.2 FC
Spark PLUS UV395™ anti-mouse I-A/I-E M5/114.15.2 FC
Brilliant Violet 750™ anti-mouse I-A/I-E M5/114.15.2 FC
Spark YG™ 593 anti-mouse I-A/I-E (Flexi-Fluor™) Antibody M5/114.15.2 FC
Spark NIR™ 685 anti-mouse I-A/I-E (Flexi-Fluor™) Antibody M5/114.15.2 FC
StarBright UltraViolet 795 anti-mouse I-A/I-E M5/114.15.2 FC
Spark PLUS B574™ anti-mouse I-A/I-E Antibody M5/114.15.2 FC
Go To Top Version: 6    Revision Date: 04/21/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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