United Kingdom (UK)
Recombinant monoclonal antibody to CD43 . Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma OX-58.
UniProt Accession Number of Target Protein: P13838
Alternative Name(s) of Target: Leukosialin; Leukocyte sialoglycoprotein; Sialophorin; W3/13 antigen; OX 58; MRC OX 58
Immunogen: This antibody was prepared by immunizing BALB/c mice with rat thymocyte leukosialin and thymocytes.
Specificity: This antibody recognises rat leukosialin, a major cell-surface sialoglycoprotein of thymocytes and T lymphocytes. It is also expressed on neutrophils, bone marrow stem cells and B-lineage cells after activation. The specificity of the leukosialin epitope is largely determined by linear protein sequence, as indicated by the reaction of this antibody with the extracellular domain of leukosialian expressed in an unglycosylated form in E. coli. However, the sensitivity of this antibody to neuraminidase treatment of leukosialin indicates that glycosylation can modify epitope conformation or flexibility.
Application Notes: This antibody binds to rat leukosialin, and can be used to label thymocytes and T lymphocytes in the lymph node cell population in flow cytometry analysis. This antibody has also been used, along with other anti-CD43 MABs, in a Western blot analysis to localize leukosialin epitopes.
Antibody first published in:
Cyster et al. The dimensions of the T lymphocyte glycoprotein leukosialin and identification of linear protein epitopes that can be modified by glycosylation. EMBO J. 1991 Apr;10(4):893-902. PMID:1706994
Note on publication:
Describes the original use of this antibody, along with other anti-CD43 MAbs, in FC, WB and plate binding assay studies to determine the dimensions of the extracellular part of the leukosialin molecule, and to establish that the antigenic determinants recognized are largely based on linear protein sequences.
Flow cytometry using the Anti-CD43 antibody OX-58 (Ab00586). Paraformaldehyde fixed rat splenocytes permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of OX-58 (Ab00586-23.0, 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.
Immunofluorescence staining of fixed rat splenocytes with anti-CD43 antibody OX-58 (Ab00586). Immunofluorescence analysis of paraformaldehyde fixed rat splenocytes on Shi-fix™ cover-slips stained with the chimeric rabbit IgG version of OX-58 (Ab00586-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane and cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00586-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.