United Kingdom (UK)
Recombinant monoclonal antibody to CD8a. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma C8/144B.
UniProt Accession Number of Target Protein: P01732
Alternative Name(s) of Target: Leu2; CD8; CD8A; T-cell surface glycoprotein CD8 alpha chain; T-lymphocyte differentiation antigen T8/Leu-2; CD8a; p32
Immunogen: This antibody was raised by immunising Balb/c mice with a synthetic peptide corresponding to the 13 C-terminal residues of the cytoplasmic domain of human CD8a.
Specificity: This antibody recognises alpha chain of human CD8 (CD8a). CD8 is a cell surface receptor expressed either as a heterodimer with the CD8 beta chain (CD8 alpha/beta) or as a homodimer (CD8 alpha/alpha). CD8 is a T-cell co-receptor involved in the recognition of MHC class I molecules. It is present on cytotoxic/suppressor T lymphocytes.
Application Notes: Tis anti-CD8 antibody can be used to detect suppressor/cytotoxic T-cells and their neoplastic counterparts in tissue sections (van Gool et al., 2015). C8/144B has been extensively applied in multiple studies where the detection of CD8+ lymphocytes was needed, such as characterisation of various inflammatory diseases, cancer treatment etc. (Petukhova et al., 2010; Stover et al., 2018). For mature T-cells, presence of CD8 and CD4 is mutually exclusive, and thus using this anti-CD8 antibody with another one diected against CD4 might serve a way of distinguishing between different T-cell subpopulation. Furthermore, C8/144B antibody can be utilised in the identification of certain neoplasms when presence or absence of the anti-CD4 signal is of a diagnostic value. For example, in T-lymphoblastic lymphoma/leukemia, CD4 and CD8 are often co-expressed.
Antibody first published in:
Mason et al. Immunohistological detection of human cytotoxic/suppressor T cells using antibodies to a CD8 peptide sequence. J Clin Pathol. 1992 Dec;45(12):1084-8. PMID:1479035
Note on publication:
This article describes generation and characterisation of C8/144B antibody.
Western Blot using anti-CD8a antibody C8/144B (Ab01344). human lymph node tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric mouse IgG version of C8/144B (Ab01344-1.1) at 0.3 µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of Jurkat cells using anti-CD8a (Ab01344) C8/144B Immunofluorescence analysis of paraformaldehyde fixed Jurkat cells stained with the chimeric mouse IgG version of C8/144B (Ab01344-1.1) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01215-1.1, DAPI, merged channels and an isotype control. The isotype control was stained with anti-unknown antibody (Ab00178-1.1) followed by Alexa Fluor® 488 secondary antibody.
Flow-cytometry using the anti- antibody (Ab01344). were stained with anti-unknown IgG antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG-chimeric version of (Ab01344-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.
Western Blot using anti-CD8a antibody C8/144B (Ab01344). human lymph node tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric mouse IgG version of C8/144B (Ab01344-1.1) at 0.3 µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Flow cytometry using the Anti-CD8a antibody C8/144B (Ab01344). Paraformaldehyde fixed Jurkat cells were stained with anti-unknown specificity antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG1 version of C8/144B (Ab01344-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.