United Kingdom (UK)
Recombinant monoclonal antibody to CD53. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma HD77.
UniProt Accession Number of Target Protein: P19397
Alternative Name(s) of Target: TSPAN-25; Leukocyte surface antigen CD53; Tetraspanin-25; MOX44; TSPAN25
Immunogen: This antibody was raised by immunising mice with human leukemia cells.
Specificity: This antibody is specific for the extracytoplasmic part of human CD53, a pan-leukocyte antigen.
Application Notes: This antibody has applications in the detection of normal and neoplastic white blood cells. This antibody has been used to stain tumour and mononuclear cells for CD53 expression in flow cytometric analysis of samples from ovarian carcinoma patients (Marme et al, 2002), and cells of the colon carcinoma line HT29 (Koretz et al, 1992). In immunohistochemical analysis of frozen tissue sections, this antibody has been shown to react with osteoclasts and osteoclast-like giant cells in malignant fibrous histiocytoma and in benign giant cell tumor of tendon sheath (Mechtersheimer et al, 1990). This antibody has been used to re-immunoprecipitate CD53 from PKCα immunoprecipiates (Zhang et al, 2001).
Antibody first published in:
Mechtersheimer et al. Expression of HLA-A,B,C, beta 2-microglobulin (beta 2m), HLA-DR, -DP, -DQ and of HLA-D-associated invariant chain (Ii) in soft-tissue tumors. Int J Cancer. 1990 Nov 15;46(5):813-23. PMID:2228310
Note on publication:
Describes the original use of this antibody, and its use in immunohistochemical analysis of frozen tissue sections.
Immunofluorescence staining of fixed Daudi cells with anti-CD53 antibody HD77 (Ab01428) Immunofluorescence analysis of paraformaldehyde fixed Daudi cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version HD77 (Ab01428-23.0) at 10 µg/ml for 1h 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 Ab01428-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-1.1) followed by staining with Alexa Fluor® 488 secondary antibody.
Western Blot using anti-CD53 antibody HD77 (Ab01428) Human lymph node (A), human thymus (B) and human spleen (C) tissue lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of HD77 (Ab01428-23.0) at 0.1 µg/ml, 0.3 µg/ml and 0.5 µg/ml, respectively, before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of fixed Daudi cells with anti-CD53 antibody HD77 (Ab01428) Immunofluorescence analysis of paraformaldehyde fixed Daudi cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version HD77 (Ab01428-23.0) at 10 µg/ml for 1h 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 Ab01428-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-1.1) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow cytometry using the Anti-CD53 antibody HD77 (Ab01428). Paraformaldehyde fixed Daudi cells were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of HD77 (Ab01428-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.