CD8 Antibody (53-6.7) [HRP]

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-49045H

Novus Biologicals, part of Bio-Techne
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Key Product Details

Species Reactivity

Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Cell depletion, Inhibition of T Cell Function, CyTOF-ready

Label

HRP

Antibody Source

Monoclonal Rat IgG2a Kappa Clone # 53-6.7

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all CD8 Antibodies »

Product Specifications

Immunogen

Mouse thymus or spleen.

Localization

Most thymocytes, T cell subset, some NK cells

Clonality

Monoclonal

Host

Rat

Isotype

IgG2a Kappa

Theoretical MW

27 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Applications for CD8 Antibody (53-6.7) [HRP]

Application
Recommended Usage

Cell depletion

Optimal dilutions of this antibody should be experimentally determined.

CyTOF-ready

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry-Frozen

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry-Paraffin

Optimal dilutions of this antibody should be experimentally determined.

Immunoprecipitation

Optimal dilutions of this antibody should be experimentally determined.

Inhibition of T Cell Function

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Each lot of this CD8a antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is <0.25ug/10^6 cells in 100ul volume. It is recommended that the reagent be titrated for optimal performance for each application. 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: immunoprecipitation, in vivo and in vitro cell depletion, inhibition of CD8 T cell proliferation, blocking of cytotoxicity, and immunohistochemical staining of both acetone-fixed frozen sections and zinc-fixed paraffin-embedded sections. This antibody is not recommended for immunohistochemistry of formalin-fixed paraffin sections. Immunocytochemistry/Immunofluorescence was reported in scientific literature. Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID 24565643)

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Preservative

No Preservative

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C in the dark.

Background: CD8

CD8, also known as Leu-2 or T8 in human and Lyt2 or Lyt3 in mouse, is a cell surface glycoprotein belonging to the immunoglobulin supergene family (1, 2). CD8 is expressed on cytotoxic T-lymphocytes (T-cells), most thymocytes, between 35-45% of peripheral blood lymphocytes, and a population of natural killer (NK) cells (1, 2). The CD8 molecule consists of disulfide-linked alpha (alpha) and beta (beta) chains that present on T-cells as either CD8alphaalpha homodimers or CD8alphabeta heterodimers (1, 3). Both alpha and beta chains consist of a signaling sequence, an extracellular Ig-like domain, a membrane proximal stalk region, a transmembrane domain, and a cytoplasmic tail (3). Human CD8alpha is processed as 235 amino acids (aa) in length with a theoretical molecular weight of ~26 kDa, while mouse CD8alpha is 247 aa and has a theoretical molecular weight of 27.5 kDa (4, 5). Functionally, CD8 acts as an antigen coreceptor on cytotoxic T-cells and interacts with the major histocompatibility complex (MHC) class I molecules on antigen presenting cells (APCs), mediating cell-cell interactions within the immune system. Conversely, CD4 molecules interact with antigens presented on MHC class II molecules and are activated to become helper T-cells (TH) (1,2). Interestingly, thymocytes can transiently express both CD4 and CD8 during the maturation process (2). Furthermore, the cytoplasmic tail of CD8 has a Lck (lymphocyte-specific protein tyrosine kinase) binding domain where Lck interacts with CD8, initiating a phosphorylation cascade that activates transcription factors and promotes T-cell activation (6). More specifically, CD8alphabeta functions as a T-cell co-receptor, while CD8alphaalpha promotes T-cell survival and differentiation (7).

Given its role in the immune system, CD8-deficiency in T-cells is a hallmark of many diseases and pathologies (8-10). Specifically, CD8+ T-cell deficiency is prevalent in chronic autoimmune diseases including multiple sclerosis, rheumatoid arthritis, ulcerative colitis, Crohn's disease, type 1 diabetes mellitus, and Graves' disease (8). Furthermore, cancers or chronic infection can lead to CD8 T-cell exhaustion as the continual antigen presentation and inflammatory signals eventually cause the CD8+ T-cells to lose functionality (9, 10). However, animal models and clinical studies have suggested that T-cells are capable of being reinvigorated using inhibitory receptor blockade resulting in better disease outcomes and these exhausted T-cells may be a potential therapeutic target (9, 10).

Alternative names for CD8 includes CD antigen: CD8a, CD8 antigen, alpha polypeptide (p32), CD8a molecule, CD8A, Leu2 T-lymphocyte antigen, LEU2, MAL, OKT8 T-cell antigen, p32, T cell co-receptor, T8 T-cell antigen, T-cell antigen Leu2, T-cell surface glycoprotein CD8 alpha chain, and T-lymphocyte differentiation antigen T8/Leu-2.

References

1. Littman D. R. (1987). The structure of the CD4 and CD8 genes. Annual review of immunology. https://doi.org/10.1146/annurev.iy.05.040187.003021

2. Naeim F. (2008). Chapter 2- Principles of Immunophenotyping. Hematopathology. https://doi.org/10.1016/B978-0-12-370607-2.00002-8.

3. Gao, G. F., & Jakobsen, B. K. (2000). Molecular interactions of coreceptor CD8 and MHC class I: the molecular basis for functional coordination with the T-cell receptor. Immunology today. https://doi.org/10.1016/s0167-5699(00)01750-3

4. UniProt (P01732)

5. UniProt (P01731)

6. Kappes D. J. (2007). CD4 and CD8: hogging all the Lck. Immunity. https://doi.org/10.1016/j.immuni.2007.11.002

7. Gangadharan, D., & Cheroutre, H. (2004). The CD8 isoform CD8alphaalpha is not a functional homologue of the TCR co-receptor CD8alphabeta. Current opinion in immunology. https://doi.org/10.1016/j.coi.2004.03.015

8. Pender M. P. (2012). CD8+ T-Cell Deficiency, Epstein-Barr Virus Infection, Vitamin D Deficiency, and Steps to Autoimmunity: A Unifying Hypothesis. Autoimmune diseases. https://doi.org/10.1155/2012/189096

9. Kurachi M. (2019). CD8+ T cell exhaustion. Seminars in immunopathology. https://doi.org/10.1007/s00281-019-00744-5

10. Hashimoto, M., Kamphorst, A. O., Im, S. J., Kissick, H. T., Pillai, R. N., Ramalingam, S. S., Araki, K., & Ahmed, R. (2018). CD8 T Cell Exhaustion in Chronic Infection and Cancer: Opportunities for Interventions. Annual review of medicine. https://doi.org/10.1146/annurev-med-012017-043208

Alternate Names

CD8, CD8A

Gene Symbol

CD8A

Additional CD8 Products

Product Documents for CD8 Antibody (53-6.7) [HRP]

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for CD8 Antibody (53-6.7) [HRP]

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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