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Spark YG™ 593 anti-mouse I-A/I-E (Flexi-Fluor™) 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
Compare all formats See Spark YG™ 593 spectral data
Cat # Size Price Quantity Save
285446 50 µg 269€
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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 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
  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

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 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
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
Go To Top Version: 1    Revision Date: 06.02.2025

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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