Western blot analysis of AFP monoclonal antibody, clone AF5 (Cat # RAB01092). HepG2 cell lysate (35 ug protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit antiby (Cat # RAB01092) at 1 ug/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). Cat # RAB01092 successfully detected human AFP in HepG2 cell lysates.
Immunofluorescence
Immunofluoresence staining of fixed HepG2 cells with AFP monoclonal antibody, clone AF5 (Cat # RAB01092). Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells, permeabilized with 0.15% Triton stained with the chimeric rabbit antibody (Cat # RAB01092) at 10 ug/mL for 1h followed by Alexa Fluor®488 secondary antibody (1 ug/mL), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Cat # RAB01092, DAPI, merged channels and a negative control. The negative control was stained with unimmunized rabbit IgG followed by Alexa Fluor®488 secondary antibody.
Flow Cytometry
Flow-cytometry using AFP monoclonal antibody, clone AF5 (Cat # RAB01092) HepG2 cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric antibody (Cat # RAB01092, blue line) at a concentration of 10 ug/mL for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody at 2 ug/mL and cells analyzed on a FACSCanto flow-cytometer.
This gene encodes alpha-fetoprotein, a major plasma protein produced by the yolk sac and the liver during fetal life. Alpha-fetoprotein expression in adults is often associated with hepatoma or teratoma. However, hereditary persistance of alpha-fetoprotein may also be found in individuals with no obvious pathology. The protein is thought to be the fetal counterpart of serum albumin, and the alpha-fetoprotein and albumin genes are present in tandem in the same transcriptional orientation on chromosome 4. Alpha-fetoprotein is found in monomeric as well as dimeric and trimeric forms, and binds copper, nickel, fatty acids and bilirubin. The level of alpha-fetoprotein in amniotic fluid is used to measure renal loss of protein to screen for spina bifida and anencephaly. [provided by RefSeq