United Kingdom (UK)
Recombinant monoclonal antibody to Alpha-fetoprotein (AFP). Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma AF5.
UniProt Accession Number of Target Protein: P02771
Alternative Name(s) of Target: A-Feto-protein
Immunogen: AF5 was prepared by immunizing female BALB/c mice with human amniotic fluid containing AFP, from a pregnancy affected by a neural tube defect, diluted and emulsified in Freund's complete adjuvant.
Specificity: Recognises the human Alpha-fetoprotein.
Antibody first published in: Hemminki et al. Introduction of lysine residues on the light chain constant domain improves the labelling properties of a recombinant Fab fragment. Protein Eng. 1995 Feb;8(2):185-91. PMID:7543205
Flow-cytometry using the anti-Alpha-fetoprotein AF5 (Ab00213) HepG2 cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of AF5 (Ab00213-23.0, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.
Immunofluoresence staining of fixed HepG2 cells with anti-Alpha-fetoprotein (AFP) antibody AF5 (Ab00213) Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells, permeabilized with 0.15% Triton stained with the chimeric rabbit IgG version of AF5 (Ab00213-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (1 µg/ml), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00213-23.0, DAPI, merged channels and a negative control. The negative control was stained with unimmunized rabbit IgG followed by Alexa Fluor® 488 secondary antibody.
Western Blot using anti-Alpha-fetoprotein antibody AF5 (Ab00213) HepG2 cell lysate (35µg protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of AF5 (Ab00213-23.0) at 1 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 Alpha-fetoprotein is 68.7kDa, though this protein is also glycosylated (Uniprot ID: P02771). Ab00213-23.0 successfully detected human Alpha-fetoprotein in HepG2 cell lysates.
Flow-cytometry using the anti-Alpha-fetoprotein AF5 (Ab00213) HepG2 cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of AF5 (Ab00213-23.0, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.