United Kingdom (UK)
Recombinant monoclonal antibody to CD45RA or A/B. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma OX-30.
UniProt Accession Number of Target Protein: P04157
Alternative Name(s) of Target: PTPRC; B220; CD45; CD45 R; GP180; L-CA; LCA; LY5; T200; protein tyrosine phosphatase; receptor type C; Receptor-type tyrosine-protein phosphatase C
Immunogen: This antibody was prepared by immunizing BALB/c mice with rat lymph node glycoproteins.
Specificity: This antibody recognizes all molecular forms of rat L-CA, and is noncompetitive with OX-1, 28 and 29 in binding to L-CA. While purified L-CA inhibited binding of this antibody to lymph node cells, no such inhibition of binding was seen with reduced and alkylated L-CA.
Application Notes: This antibody binds all molecular forms of rat L-CA, and consequently can be used to label thymocytes, T cells, B cells and most bone marrow cells (Woollett, 1985).
Antibody first published in:
Woollett et al. Molecular and antigenic heterogeneity of the rat leukocyte-common antigen from thymocytes and T and B lymphocytes. Eur J Immunol. 1985 Feb;15(2):168-73. PMID:2578966
Note on publication:
Describes the original use of this antibody in FC and blocking analysis to assess the molecular and antigenic heterogenity of rat L-CA.
Flow-cytometry using anti-CD45RA or A/B (Receptor-type tyrosine-protein phosphatase C isoform) antibody OX-30 (Ab00576) Staining of rat (Rattus norvegicus) lymphocytes with either an isotype control (A) or the rabbit-chimeric version of OX-30 (B, Ab00576-23.0) at a concentration of 1 µg/ml for 30 mins at RT. After washing, bound antibody was detected using an Alexa Fluor ® 488 conjugated donkey anti-rabbit antibody (ab150073) and cells analysed on a FACSCanto flow-cytometer.
Immunofluorescence staining of fixed rat splenocytes with anti-CD45RA or A/B (Receptor-type tyrosine-protein phosphatase C) antibody OX-30 (Ab00576) Immunofluorescence analysis of paraformaldehyde fixed rat (Rattus norvegicus) splenocytes on Shi-fix™ coverslips, permeabilized with 0.15% Triton stained with the chimeric rabbit version of OX-30 (Ab00576-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (1 µg/ml), showing membrane and cytoplasmic staining of a subset of cells. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00576-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with an anti-Fluorescein antibody (Ab102-23.0) followed by Alexa Fluor® 488 secondary antibody.
Western Blot using anti-CD45RA or A/B (Receptor-type tyrosine-protein phosphatase C) antibody OX-30 (Ab00576) Rat thymus lysate samples (35µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of OX-30 (Ab00576-23.0) at 2 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for unmodified CD45RA is ~143.3kDa, but this protein is also highly glycosylated (UniProt Id: P04157). Ab00576-23.0 successfully detected CD45RA or A/B in rat thymus tissue lysate.