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Safety Data Sheet (SDS) Brilliant Violet 421 Anti-mouse I-A/I-E Antibody     Product Data Sheet (PDF)    
Brilliant Violet 421™ Anti-mouse I-A/I-E Antibody
1138155 125 µl $155.00       
1138160 50 µg $225.00       
Clone: M5/114.15.2
Isotype: Rat IgG2b, κ
Reactivity: Mouse
Immunogen: Activated C57BL/6 mouse spleen cells
Formulation: µl size: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
µg size: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation: The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 421™ under optimal conditions. The solution is free of unconjugated Brilliant Violet 421™ and unconjugated antibody.
Concentration: µl size: lot-specific (please contact technical support for concentration and total µg amount)
µg size: 0.2 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
IHC - Validated

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, and in vitro and in vivo blocking of antigen presentation or ligand binding4-7.

Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining using the µl size, the suggested use of this reagent is ≤5 µl per million cells or 5 µl per 100 µl of whole blood. For flow cytometric staining using the µg size, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.

Application References:

1. Bhattacharya A, et al. 1981. J. Immunol. 127:2488. (IP)
2. Viville S, et al. 1993. Cell 72:635. (IHC)
3. Nelson AJ, et al. 1993. J. Immunol. 151:2453. (IHC)
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.
10. Christensen SR, et al. 2006. Immunity 25:417. PubMed
11. Matte-Martone C, et al. 2008. Blood 111:3884. PubMed
12. De Pascalis R, et al. 2008. Infect. Immun. 76:4311. PubMed
13. Kuns RD, et al. 2009. Blood 113:5999. PubMed
14. Sabatino JJ, et al. 2011. J. Exp. Med. 208:81. PubMed
15. Draber P, et al. 2011. Mol Cell Biol. 22:4550. PubMed
16. Fu H, et al. 2014. Nat Commun. 5:3436. PubMed

C57BL/6 mouse splenocytes were stained

C57BL/6 mouse splenocytes were stained with mouse I-A/I-E (clone M5/114.15.2) Brilliant Violet 421™ (filled histogram) or rat IgG2b, κ Brilliant Violet 421™ isotype control (open histogram).

BL/6 mouse lymph nodes, fixed

BL/6 mouse lymph nodes, fixed O/N in PLP, blocked with 10% rat serum, stained with I-A/I-E BV421™ (red), B220 Alexa Fluor® 647 (blue), and CD11c FITC (green) in 1% BSA and 0.1% Tween-20 in PBS. Images were acquired with an automated widefield microscope (Nikon Eclipse Ti) and a CCD camera (QImaging Retiga 2000R). Emitted light was collected through 440/40, 525/50, and 700/75 nm bandpass filters. Images provided by Ann Haberman and Christine Podolski, Yale University.



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.

Other Names: MHC class II
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
Antigen References:

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