- 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
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 DetailsProduct 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 Spark YG™ 593 under optimal conditions.
- Concentration
- 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
- Recommended Usage
-
Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.
For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
* Spark YG™ 593 has a maximum excitation of 573 nm and a maximum emission of 593 nm. - Excitation Laser
-
Green Laser (532 nm)/Yellow-Green Laser (561 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
-
For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.
-
Application References
(PubMed link indicates BioLegend citation) -
- Bhattacharya A, et al. 1981. J. Immunol. 127:2488. (IP)
- Viville S, et al. 1993. Cell 72:635. (IHC)
- Nelson AJ, et al. 1993. J. Immunol. 151:2453. (IHC)
- Shi Y, et al. 1998. J. Exp. Med. 187:367. (Block)
- Yamashita I, et al. 1993. Int. Immunol. 5:1139.
- Guo M, et al. 1995. Zygote 3:65. (IHC)
- Kim A, et al. 2004. Exp. Mol. Med. 36:428. (Block)
- Luckashenak NA, et al. 2006. J. Immunol. 177:5177.
- Venanzi ES, et al. 2007. J. Immunol. 179:5693.
- Christensen SR, et al. 2006. Immunity 25:417. PubMed
- Matte-Martone C, et al. 2008. Blood 111:3884. PubMed
- De Pascalis R, et al. 2008. Infect. Immun. 76:4311. PubMed
- Kuns RD, et al. 2009. Blood 113:5999. PubMed
- Sabatino JJ, et al. 2011. J. Exp. Med. 208:81. PubMed
- Draber P, et al. 2011. Mol Cell Biol. 22:4550. PubMed
- Fu H, et al. 2014. Nat Commun. 5:3436. PubMed
- Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
- Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
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
- What are Flexi-Fluors?
-
Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer.
- How quickly will I receive my order?
-
We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary.
- How are Flexi-Fluors different from regular catalog products?
-
Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.
- How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?
-
Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL.
For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
- I can’t find the antibody-dye combination that I need. When will it be available?
-
We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services.
- I need help to validate the performance of my Flexi-Fluor. Who should I contact?
-
Please get in touch with Technical Service for assistance.
- Can I order more than 50 μg of a Flexi-Fluor?
-
Yes, you can order multiple vials of the same Flexi-Fluor products. We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.
- What is the expiration date of my Flexi-Fluor?
-
Expiration dates can be found on the vial label or by using our CoA lookup tool.
Other Formats
View All I-A/I-E Reagents Request Custom ConjugationCompare Data Across All Formats
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.
-
Biotin anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
FITC anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
PE anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
Purified anti-mouse I-A/I-E
C57/B6 mouse splenocytes were stained with purified anti-mou...
Fresh, frozen mouse spleen was stained with purified I-A/I-E... -
PE/Cyanine5 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
APC anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
Alexa Fluor® 488 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I...
Paraformaldehyde-fixed (1%), 500 µm-thick mouse spleen secti... -
Alexa Fluor® 647 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
Pacific Blue™ anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
Alexa Fluor® 700 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I...
Confocal image of C57BL/6 mouse spleen sample acquired using...
Confocal image of C57BL/6 mouse thymus sample acquired using...
Confocal image of C57BL/6 mouse lung sample acquired using t...
Confocal image of C57BL/6 mouse small intestine sample acqui...
Confocal image of C57BL/6 mouse liver sample acquired using ... -
PerCP/Cyanine5.5 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with CD45R/B220 APC a... -
PerCP anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
APC/Cyanine7 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD11b... -
PE/Cyanine7 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse I-A/I... -
Brilliant Violet 421™ anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with mouse I-A/I-E (c...
BL/6 mouse lymph nodes, fixed O/N in PLP, blocked with 10% r...
Mice were injected subcutaneously with sheep red blood cells... -
Brilliant Violet 510™ anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with mouse I-A/I-E (c... -
Purified anti-mouse I-A/I-E (Maxpar® Ready)
C57BL/6 mouse bone marrow cells stained with 174Yb-anti-I-A/... -
Brilliant Violet 605™ anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with mouse I-A/I-E (c... -
Brilliant Violet 650™ anti-mouse I-A/I-E
C57 mouse splenocytes were stained with I-A/I-E (clone M5/11... -
Brilliant Violet 711™ anti-mouse I-A/I-E
C57 mouse splenocytes were stained with I-A/I-E (clone M5/11... -
Brilliant Violet 785™ anti-mouse I-A/I-E
C57 mouse splenocytes were stained with I-A/I-E (clone M5/11... -
PE/Dazzle™ 594 anti-mouse I-A/I-E
C57 mouse splenocytes were stained with I-A/I-E (clone M5/11... -
Alexa Fluor® 594 anti-mouse I-A/I-E
C57BL/6 mouse frozen spleen section was fixed with 4% parafo...
Paraformaldehyde-fixed (4%), 500 μm-thick mouse spleen secti... -
APC/Fire™ 750 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with mouse I-A/I-E (c... -
TotalSeq™-A0117 anti-mouse I-A/I-E
-
Ultra-LEAF™ Purified anti-mouse I-A/I-E
C57/B6 mouse splenocytes were stained with LEAF™ anti-mouse ... -
TotalSeq™-B0117 anti-mouse I-A/I-E
-
TotalSeq™-C0117 anti-mouse I-A/I-E
-
Spark Blue™ 550 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes stained with I-A/I-E (clone M5/114... -
PE/Fire™ 640 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse/human... -
Spark YG™ 581 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD45R... -
PE/Fire™ 810 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD45R... -
Spark UV™ 387 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD45R... -
Spark Violet™ 538 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD45... -
PerCP/Fire™ 806 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD45R... -
Spark Red™ 718 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse/human... -
APC/Fire™ 810 anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse/human... -
Spark PLUS UV395™ anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD45R... -
Brilliant Violet 750™ anti-mouse I-A/I-E
C57BL/6 mouse splenocytes were stained with anti-mouse CD45R... -
Spark YG™ 593 anti-mouse I-A/I-E (Flexi-Fluor™) Antibody
-
Spark NIR™ 685 anti-mouse I-A/I-E (Flexi-Fluor™) Antibody
Login / Register 



Follow Us