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Alexa Fluor® 594 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
A_M5-114-15-2_A594_I-AI-E_1_Antibody_061019
C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS plus 5% rat/mouse serum for 30 minutes at room temperature. Then the section was stained with 2.5 µg/ml of I-A/I-E (clone M5/114.15.2) Alexa Fluor® 594 (red), 5 µg/ml of F4/80 (clone BM8) Alexa Fluor® 647 (blue), and 5 µg/ml of CD3 (clone 17A2) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured with a 10X objective.
  • A_M5-114-15-2_A594_I-AI-E_1_Antibody_061019
    C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS plus 5% rat/mouse serum for 30 minutes at room temperature. Then the section was stained with 2.5 µg/ml of I-A/I-E (clone M5/114.15.2) Alexa Fluor® 594 (red), 5 µg/ml of F4/80 (clone BM8) Alexa Fluor® 647 (blue), and 5 µg/ml of CD3 (clone 17A2) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured with a 10X objective.
  • B_M5slash114dot15dot2_A594_I-A_I-E_Antibody_3D-IHC_092121.png
    Paraformaldehyde-fixed (4%), 500 μm-thick mouse spleen section was processed according to the Ce3DTM Tissue Clearing Kit protocol (cat. no. 427701). The section was costained with anti-mouse I-A/I-E Antibody (clone M5/114.15.2) Alexa Fluor® 594 at 5 µg/mL (green), and anti-mouse CD146 Antibody (Clone ME-9F1) Alexa Fluor® 647 at 5 µg/mL (magenta). The section was then optically cleared and mounted in a sample chamber. The image was captured with a 10X objective using Zeiss 780 confocal microscope and processed by Imaris image analysis software.
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107650 100 µg $253
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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

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® 594 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

IHC-F - Quality tested
3D IHC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by immunohistochemical staining on frozen tissue sections. For immunohistochemistry, a concentration range of 0.1 - 10 μg/ml is recommended. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


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

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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).

Application References

(PubMed link indicates BioLegend citation)
  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
  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
  1. Nishida Y, et al. 2023. Sci Adv. eadh1436:9. PubMed
  2. Dixit S, et al. 2018. Biomaterials. 159:130. PubMed
RRID
AB_2566438 (BioLegend Cat. No. 107650)

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

There are no FAQs for this product.

Other Formats

View All I-A/I-E Reagents Request Custom Conjugation
Description Clone Applications
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
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
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
TotalSeq™-B0117 anti-mouse I-A/I-E M5/114.15.2 PG
TotalSeq™-C0117 anti-mouse I-A/I-E M5/114.15.2 PG
Spark Blue™ 550 anti-mouse I-A/I-E M5/114.15.2 FC
PE/Fire™ 640 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
Go To Top Version: 2    Revision Date: 09/21/2021

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.

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