alpha-Fetoprotein/AFP Antibody (AF5) - Chimeric - Azide and BSA Free

Western Blot: alpha-Fetoprotein/AFP Antibody (AF5) [NBP2-82221] - HepG2 cell lysate (35ug protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of alpha-Fetoprotein/AFP Antibody (AF5) [NBP2-82221] 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). NBP2-82221 successfully detected human Alpha-fetoprotein in HepG2 cell lysates.

Immunocytochemistry/Immunofluorescence: alpha-Fetoprotein/AFP Antibody (AF5) [NBP2-82221] - Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells, permeabilized with 0.15% Triton stained with the chimeric rabbit IgG version of alpha-Fetoprotein/AFP Antibody (AF5) [NBP2-82221] 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: NBP2-82221, 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: alpha-Fetoprotein/AFP Antibody (AF5) [NBP2-82221] - HepG2 cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of ([NBP2-82221], blue line) at a concentration of 10 ug/ml for 30 mins at RT. After washing, bound antibody was detected using an anti-rabbit IgG JK (FITC-conjugate) antibody at 2 ug/ml and cells analyzed on a FACSCanto flow-cytometer.

• Reactivity: Hu
• Applications: WB, Flow, ICC/IF
• Clone: AF5
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1 mg/ml

alpha-Fetoprotein/AFP Antibody (AF5) - Chimeric - Azide and BSA Free Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: AF5 was prepared by immunizing female BALB/c mice with human amniotic fluid containing alpha-Fetoprotein/AFP, from a pregnancy affected by a neural tube defect, diluted and emulsified in Freund's complete adjuvant.
• Specificity: Recognises the human alpha-Fetoprotein/AFP.
• Isotype: IgG Kappa
• Clonality: Monoclonal
• Host: Rabbit
• Gene: AFP
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Immunocytochemistry/ Immunofluorescence
  • Western Blot
• Application Notes: This chimeric rabbit antibody was made using the variable domain sequences of the original murine IgG1 format, for improved compatibility with existing reagents, assays and techniques. 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

Packaging, Storage & Formulations

• Storage: Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.
• Buffer: PBS
• Preservative: 0.02% Proclin 300
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for alpha-Fetoprotein/AFP Antibody (AF5) - Chimeric - Azide and BSA Free

• AFP
• Alpha-1-fetoprotein
• alpha-fetoglobulin
• alpha-Fetoprotein
• DSCAM2
• FETA
• HP
• HPAFP

Background

Alpha 1 Fetoprotein 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: AFP
• Uniprot:
  • P02771(Human)