Alexa Fluor® 594 anti-mouse CD8a 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
53-67_A594_CD8a_Antibody_1072414
C57BL/6 mouse frozen lymph node section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS plus 5% rat serum for 1 hour at room temperature. Then the section was stained with 2.5 µg/ml of CD8 (clone 53-6.7) Alexa Fluor® 594 (red), 2.5 µg/ml of CD4 (clone GK1.5) Alexa Fluor® 647 (green), and 2.5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (blue) overnight at 4°C. The image was captured by 10X objective.
  • 53-67_A594_CD8a_Antibody_1072414
    C57BL/6 mouse frozen lymph node section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS plus 5% rat serum for 1 hour at room temperature. Then the section was stained with 2.5 µg/ml of CD8 (clone 53-6.7) Alexa Fluor® 594 (red), 2.5 µg/ml of CD4 (clone GK1.5) Alexa Fluor® 647 (green), and 2.5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (blue) overnight at 4°C. The image was captured by 10X objective.
  • 53-67_A594_CD8a_Antibody_2_112520
    C57BL/6 mouse frozen lymph node section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS plus 5% rat serum for 1 hour at room temperature. Then the section was stained with 2.5 µg/ml of CD8 (clone 53-6.7) Alexa Fluor® 594 (red), 2.5 µg/ml of CD4 (clone GK1.5) Alexa Fluor® 647 (green), and 2.5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (blue) overnight at 4°C. The image was captured by 10X objective.
  • 53-67_A594_CD8a_Antibody_3_092121.png
    Paraformaldehyde-fixed (1%), 500 µm-thick mouse thymus section was processed according to the Ce3D™ Tissue Clearing Kit protocol (Cat. No. 427701). The section was costained with anti-mouse CD68 Antibody (clone FA-11) Alexa Fluor® 488 at 5 µg/mL (green), anti-mouse CD8a Antibody (clone 53-6.7) Alexa Fluor® 594 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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100758 100 µg $206
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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

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 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
FC, 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 2.0 - 5.0 μg/mL is suggested. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. 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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Excitation Laser
Green Laser (532 nm)/Yellow-Green Laser (561 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).

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
Product Citations
  1. Kawabe T, et al. 2020. Nat Commun. 2.795833333. PubMed
  2. Hirai T, et al. 2020. Immunity. 54(1):84-98.e5. PubMed
  3. He R, et al. 2021. J Immunol. 206:2353. PubMed
  4. Baptista AP et al. 2019. Immunity. 50(5):1188-1201 . PubMed
  5. Mogilenko DA, et al. 2020. Immunity. 54(1):99-115.e12. PubMed
  6. Devalaraja S, et al. 2020. Cell. 1098:180. PubMed
  7. Dodagatta-Marri E, et al. 2021. Cell Rep. 36:109309. PubMed
  8. Carozza JA, et al. 2020. Nat Cancer. 184:1. PubMed
  9. An S, et al. 2019. Small. 15:e1805182. PubMed
  10. Chung YM, et al. 2021. J Immunother Cancer. 9:. PubMed
  11. Dang MT, et al. 2021. Cell Reports. 34(13):108917. PubMed
  12. Li H, et al. 2021. Nat Commun. 12:7149. PubMed
  13. Green WD, et al. 2022. J Leukoc Biol. 111:147. PubMed
  14. Tao Z, et al. 2019. Clin Cancer Res. 25:1113. PubMed
  15. Li Z, et al. 2022. Nat Commun. 13:6321. PubMed
  16. Li B, et al. 2018. Cancer Res. 78:3938. PubMed
  17. Liu P, et al. 2020. Mol Cell. 748:77. PubMed
  18. Agudo J et al. 2018. Immunity. 48(2):271-285 . PubMed
  19. Shu L, et al. 2020. Clinical Science. 134(13):1679-1696. PubMed
RRID
AB_2563237 (BioLegend Cat. No. 100758)

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

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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 UV395™ anti-mouse CD8a 53-6.7 FC
PerCP/Fire™ 780 anti-mouse CD8a 53-6.7 FC
Go To Top Version: 5    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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