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Spark Red™ 718 anti-mouse CD8a (Flexi-Fluor™) Antibody

Pricing & Availability
Clone
53-6.7 (See other available formats)
Regulatory Status
RUO
Other Names
T8, Lyt2, Ly-2
Isotype
Rat IgG2a, κ
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Product Citations
publications
Compare all formats See Spark Red™ 718 spectral data
Cat # Size Price Quantity Save
285050 50 µg £197
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Description

CD8, also known as Lyt-2, Ly-2, or T8, consists of disulfide-linked α and β chains that form the α(CD8a)/β(CD8b) heterodimer and α/α homodimer. CD8a is a 34 kD protein that belongs to the immunoglobulin family. The CD8 α/β heterodimer is expressed on the surface of most thymocytes and a subset of mature TCR α/β T cells. CD8 expression on mature T cells is non-overlapping with CD4. The CD8 α/α homodimer is expressed on a subset of γ/δ TCR-bearing T cells, NK cells, intestinal intraepithelial lymphocytes, and lymphoid dendritic cells. CD8 is an antigen co-receptor on T cells that interacts with MHC class I on antigen-presenting cells or epithelial cells. CD8 promotes T cell activation through its association with the TCR complex and protein tyrosine kinase lck.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Mouse thymus or spleen
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography and conjugated with Spark Red™ 718 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.
Excitation Laser
Red Laser (633 nm)
Application Notes

Clone 53-6.7 antibody competes with clone 5H10-1 antibody for binding to thymocytes3. The 53-6.7 antibody has been reported to block antigen presentation via MHC class I and inhibit T cell responses to IL-2. This antibody has also been used for depletion of CD8a+ cells. Additional reported applications (for the relevant formats) include: immunoprecipitation1,3, in vivo and in vitro cell depletion2,10,15, inhibition of CD8 T cell proliferation3, blocking of cytotoxicity3,4, immunohistochemical staining5,6 of acetone-fixed frozen sections and zinc-fixed paraffin-embedded sections, and spatial biology (IBEX)29,30. Clone 53-6.7 is not recommended for immunohistochemistry of formalin-fixed paraffin sections. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays or in vivo studies (Cat No. 100746).

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)
  1. Ledbetter JA, et al. 1979. Immunol. Rev. 47:63. (IHC, IP)
  2. Hathcock KS. 1991. Current Protocols in Immunology. 3.4.1. (Deplete)
  3. Takahashi K, et al. 1992. P. Natl. Acad. Sci. USA 89:5557. (Block, IP)
  4. Ledbetter JA, et al. 1981. J. Exp. Med. 153:1503. (Block)
  5. Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
  6. Fan WY, et al. 2001. Exp. Biol. Med. 226:1045. (IHC)
  7. Shih FF, et al. 2006. J. Immunol. 176:3438. (FC)
  8. Kamimura D, et al. 2006. J. Immunol. 177:306.
  9. Bouwer HGA, et al. 2006. P. Natl. Acad. Sci. USA 103:5102. (FC, Deplete)
  10. Kao C, et al. 2005. Int. Immunol. 17:1607. PubMed
  11. Ko SY, et al. 2005. J. Immunol. 175:3309. (FC) PubMed
  12. Rasmussen JW, et al. 2006. Infect. Immun. 74:6590. PubMed
  13. Lee CH, et al. 2009. Clin. Cancer Res. PubMed
  14. Geiben-Lynn R, et al. 2008. Blood 112:4585. (Deplete) PubMed
  15. Kingeter LM, et al. 2008. J. Immunol. 181:6244. PubMed
  16. Guo Y, et al. 2008. Blood 112:480. PubMed
  17. Andrews DM, et al. 2008. J. Virol. 82:4931. PubMed
  18. Britschqui MR, et al. 2008. J. Immunol. 181:7681. PubMed
  19. Kenna TJ, et al. 2008. Blood 111:2091. PubMed
  20. Jordan JM, et al. 2008. Infect. Immun. 76:3717. PubMed
  21. Todd DJ, et al. 2009. J. Exp. Med. 206:2151. PubMed
  22. Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed
  23. Medyouf H, et al. 2010. Blood 115:1175. PubMed
  24. Riedl P, et al. 2009. J. Immunol. 183:370. PubMed
  25. Apte SH, et al. 2010. J. Immunol. 185:998. PubMed
  26. Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed
  27. del Rio ML, et al. 2011. Transpl. Int. 24:501. (FC) PubMed
  28. Cui L, et al. 2015. J Control Release. 206:220. PubMed
  29. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  30. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed

Antigen Details

Structure
Ig superfamily, CD8α chain, 34 kD
Distribution

Most thymocytes, T cell subset, some NK cells, lymphoid dendritic cells

Function
Co-receptor for TCR
Ligand/Receptor
MHC class I molecule
Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Zamoyska R. 1994. Immunity 1:243.
3. Ellmeier W, et al. 1999. Annu. Rev. Immunol. 17:523.

Gene ID
12525 View all products for this Gene ID
UniProt
View information about CD8alpha 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 CD8a Reagents Request Custom Conjugation
Description Clone Applications
APC anti-mouse CD8a 53-6.7 FC
Biotin anti-mouse CD8a 53-6.7 FC,IHC
FITC anti-mouse CD8a 53-6.7 FC
PE anti-mouse CD8a 53-6.7 FC
PE/Cyanine5 anti-mouse CD8a 53-6.7 FC
Purified anti-mouse CD8a 53-6.7 FC,CyTOF®,IHC-F,IP
PE/Cyanine7 anti-mouse CD8a 53-6.7 FC
APC/Cyanine7 anti-mouse CD8a 53-6.7 FC
Alexa Fluor® 488 anti-mouse CD8a 53-6.7 FC,3D IHC
Alexa Fluor® 647 anti-mouse CD8a 53-6.7 FC,IHC-F,3D IHC,SB
Pacific Blue™ anti-mouse CD8a 53-6.7 FC
Alexa Fluor® 700 anti-mouse CD8a 53-6.7 FC
PerCP/Cyanine5.5 anti-mouse CD8a 53-6.7 FC
PerCP anti-mouse CD8a 53-6.7 FC
Brilliant Violet 421™ anti-mouse CD8a 53-6.7 FC,IHC,SB
Brilliant Violet 570™ anti-mouse CD8a 53-6.7 FC
Brilliant Violet 650™ anti-mouse CD8a 53-6.7 FC
Brilliant Violet 605™ anti-mouse CD8a 53-6.7 FC
Ultra-LEAF™ Purified anti-mouse CD8a 53-6.7 FC,CyTOF®,IHC,IP,Depletion,Block
Brilliant Violet 711™ anti-mouse CD8a 53-6.7 FC
Brilliant Violet 785™ anti-mouse CD8a 53-6.7 FC
Brilliant Violet 510™ anti-mouse CD8a 53-6.7 FC
Purified anti-mouse CD8a (Maxpar® Ready) 53-6.7 FC,CyTOF®
Alexa Fluor® 594 anti-mouse CD8a 53-6.7 IHC-F,FC,3D IHC
PE/Dazzle™ 594 anti-mouse CD8a 53-6.7 FC
APC/Fire™ 750 anti-mouse CD8a 53-6.7 FC
GoInVivo™ Purified anti-mouse CD8a 53-6.7 FC
TotalSeq™-A0002 anti-mouse CD8a 53-6.7 PG
Spark Blue™ 550 anti-mouse CD8a 53-6.7 FC
Spark NIR™ 685 anti-mouse CD8a 53-6.7 FC
TotalSeq™-C0002 anti-mouse CD8a 53-6.7 PG
TotalSeq™-B0002 anti-mouse CD8a 53-6.7 PG
Spark YG™ 570 anti-mouse CD8a 53-6.7 IHC-F
PE/Fire™ 640 anti-mouse CD8a 53-6.7 FC
PE/Fire™ 700 anti-mouse CD8a 53-6.7 FC
Spark Blue™ 574 anti-mouse CD8a Antibody 53-6.7 FC
Spark Violet™ 423 anti-mouse CD8a Antibody 53-6.7 FC
Spark UV™ 387 anti-mouse CD8a 53-6.7 FC
Spark Blue™ 515 anti-mouse CD8a 53-6.7 FC
APC/Fire™ 810 anti-mouse CD8a 53-6.7 FC
Spark Red™ 718 anti-mouse CD8a (Flexi-Fluor™) 53-6.7 FC
PE/Fire™ 810 anti-mouse CD8a 53-6.7 FC
Spark PLUS UV™ 395 anti-mouse CD8a 53-6.7 FC
Go To Top Version: 1    Revision Date: 02/07/2024

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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