United Kingdom (UK)
Recombinant monoclonal antibody to c-erbB-2. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma FWP51.
UniProt Accession Number of Target Protein: P04626
Alternative Name(s) of Target: CD340; Receptor tyrosine-protein kinase erbB-2; Neu; HER2; MLN19; Metastatic lymph node gene 19 protein; p185erbB2
Immunogen: 20x10^6 SKBR3 human breast tumour cells were injected into BALB/c mice. The hybridomas generated from the mice spleen cells were then screen for antibodies reactive to c-erbB-2 using immunofluorescence and immunoprecipitation.
Specificity: FWP51 binds to the extracellular domain of c-erbB-2.
Application Notes: Immunofluorescence and immunoprecipitation were used to confirm the specificity of FWP51 for c-erbB-2 (Harwerth et al, 1992). Flow cytometry showed that FWP51 binds the extracellular domain of c-erbB-2 (Harwerth et al, 1992). FWP51 has also been fused to the bacterial toxin, exotoxin A. The resultant fusion protein Fv(FWP51)-ETA selectively inhibited protein synthesis in cells expressing high levels of human c-erbB-2 protein, but not in cells expressing little to no human c-erbB-2 protein. Administration of FWP51 to mice impanted with human erbB-2 transformed NIH3T3 cells led to inhibition of the onset of tumour growth (Harwerth et al, 1993).
Antibody first published in:
Harwerth et al, 1992. Monoclonal antibodies against the extracellular domain of the erbB-2 receptor function as partial ligand agonists. J Biol Chem. 1992 Jul 25;267(21):15160-7. PMID:1353079
Note on publication:
Describes the original production of the antibody and its characterisation through immunofluorescence and immunoprecipitation.
Immunofluorescence staining of MCF7 cells using anti-c-erbB-2 (Ab01215) FWP51 Immunofluorescence analysis of paraformaldehyde fixed MCF7 cells stained with the chimeric mouse IgG version of FWP51 (Ab01215-1.1) 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-c-erbB-2 antibody FWP51 (Ab01215). Paraformaldehyde fixed MCF7 cells were stained with anti-unknown specificity antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG1 version of FWP51 (Ab01215-1.1, 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.