CD8 Antibody

Immunohistochemistry-Frozen: CD8 Antibody [NBP2-29475] - Mouse Ccdc88b expression in colon during DSS-induced colitis. Wild type (WT) mice were either not treated (NT) or given 3% DSS for 5 days followed by 3 days of water. Representative confocal microscopy images of tissue sections from colon of NT or DSS-treated mice at day 8, and stained for or Ccdc88b (red) and CD8 (green). Scale bars, 100 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29030607/) licensed under a CC-BY license.

Formalin-fixed, paraffin-embedded human tonsil stained with CD8 antibody at 2ug/ml at RT. HIER: Tris/EDTA, pH9.0, 45min. Secondary: HRP-polymer, 30min. DAB, 5min.

• Reactivity: Hu, Mu, Po
• Applications: Flow, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

CD8 Antibody Summary

• Description: 200 ug/ml of antibody purified from rabbit anti-serum by Protein A chromatography. Supplied in 10 mM PBS, pH 7.4 with 0.05% BSA and 0.05% sodium azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-33122)
  
  Antibody with azide - store at 2 to 8C. Antibody is stable for 24 months. Non-hazardous.
• Immunogen: Recombinant fragment from human CD8 C-terminal cytoplasmic domain of alpha chain (exact sequence is proprietary). (Uniprot: P01732)
• Localization: Cell surface
• Marker: Cytotoxic- & Suppressor T-Cell Marker
• Isotype: IgG Kappa
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CD8A
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1-2 ug/million cells
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1-2 ug/ml
• Application Notes: Use in Immunohistochemistry reported in scientific literature (PMID:33839961)
  Optimal dilution for a specific application should be determined.
  Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
• Theoretical MW: 32 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.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 29030607). Use in Porcine reported in scientific literature (PMID:33839961)

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 10 mM PBS, pH 7.4 with 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Protein A purified

Alternate Names for CD8 Antibody

• CD_antigen: CD8a
• CD8 antigen, alpha polypeptide (p32)
• CD8
• 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
• T-lymphocyte differentiation antigen T8/Leu-2

Background

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

Limitations

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.

Publications for CD8 Antibody (NBP2-29475)(8)

Publications using NBP2-29475

• Oswal N, Thangavel H, Lizardo K et al. Diets Differently Regulate Pulmonary Pathogenesis and Immune Signaling in Mice during Acute and Chronic Mycobacterium tuberculosis Infection Life (Basel, Switzerland) 2023-01-13 [PMID: 36676177] (IHC-P, Mouse)
• Fodil N, Moradin N, Leung V et al. CCDC88B is required for pathogenesis of inflammatory bowel disease. Nat Commun. 2017-10-13 [PMID: 29030607] (ICC/IF, Mouse)
• Vieyra-Garcia P Pathophysiology of cutaneous T-cell lymphoma: Understanding of neoplastic Tcells and non-malignant infiltrate in lesional skin Thesis 2016-01-01 (ICC/IF, Human)
• Nicolas AM, Pesic M, Engel E et al. Inflammatory fibroblasts mediate resistance to neoadjuvant therapy in rectal cancer Cancer cell 2022-02-14 [PMID: 35120600] (IF/IHC)
• Ji Y, Fan X, Zhang Y et al. Glycine regulates mucosal immunity and the intestinal microbial composition in weaned piglets Amino acids 2021-04-11 [PMID: 33839961] (IF/IHC, Porcine)
• Choi YW, Kim YH, Oh SY et al. Senescent Tumor Cells Build a Cytokine Shield in Colorectal Cancer Advanced Science 2021-01-04 [PMID: 33643790] (IF/IHC, Mouse)
• Mondal T, Shivange GN, Tihagam RG et al. Unexpected PD-L1 immune evasion mechanism in TNBC, ovarian, and other solid tumors by DR5 agonist antibodies EMBO molecular medicine 2021-02-15 [PMID: 33587338]
• Byun JK, Park M, Lee S et al. Inhibition of Glutamine Utilization Synergizes with Immune Checkpoint Inhibitor to Promote Antitumor Immunity J Virol 2020-11-04 [PMID: 33159855]

Review for CD8 Antibody (NBP2-29475) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer ICC Human 04/20/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: Human Tonsil tissue
• Species: Human

Comments

• Comments: Single staining of CD8 (red) on FFPE human tonsile sections.Nuclei are stained with DAPI(blue).CD8a Antibody [Unconjugated]NBP2-29475. Secondery antibody used is donkey anti-rabbit A647.

Bioinformatics

• Gene Symbol: CD8A
• Uniprot:
  • P01732(Human)