Native full-length human alpha-Fetoprotein (AFP) purified from human cord blood by hydrophobic, ion-exchange and affinity chromatography, sulphate fractionation and MonoQ-FPLC. 95 % SDS-PAGE pure single band at 70 kDa. Donors screened HBsAg / HIV / HCV negative. Supplied lyophilised in 0.1 mg vials (bulk pack 0.5 mg as cat. 858-0.5) for use as ELISA standard, immunisation antigen, oncology biomarker reference protein, and prenatal-screening assay calibration.
Specifications
Catalogue number: 858 (0.1 mg pack) / 858-0.5 (0.5 mg bulk pack)
Product name: alpha-Fetoprotein (human) — full-length native protein
Source: Human cord blood (donor pool screened HBsAg / HIV / HCV negative)
Molecular weight: ~70 kDa (single band on SDS-PAGE at 25 mg/lane load)
Gene symbol: AFP (4q11–q21)
UniProt: P02771
Aliases: a-Fetoprotein, aFP, AFP, alpha-fetoglobulin, hepatoma-associated fetoprotein
Purity: 95 % (SDS-PAGE, single band at 70 kDa)
Format: Lyophilised
Purification: Hydrophobic-interaction, ion-exchange, affinity chromatography, sulphate fractionation, MonoQ-FPLC polishing
Reconstitution
Reconstitute in sterile PBS pH 7.4 at the desired protein concentration. Centrifuge the vial briefly before opening; reconstitute by gentle inversion or low-speed pipetting (do not vortex). For ELISA coating, dilute the reconstituted protein to 1–10 µg/mL in carbonate / bicarbonate coating buffer (pH 9.6) immediately before use. For immunoassay standard-curve work, generate single-use frozen aliquots from a single reconstitution to remove between-batch standard-curve drift. For Western-blot positive-control loading, use 25–100 ng per lane. Avoid repeated freeze–thaw of the reconstituted aliquots.
Storage
Lyophilised protein: shipped at ambient temperature; store up to 2 years at -20 °C or lower.
Reconstituted protein: store up to 6 months at -70 °C.
Avoid repeated freeze–thaw cycles.
Applications
- ELISA / immunoassay standard for absolute quantification of serum AFP
- Sandwich-ELISA coating antigen for the detection and characterisation of anti-AFP antibody clones
- Immunisation antigen for the production of polyclonal and monoclonal anti-AFP antibodies (used in the generation of YO cat. 564 / cat. 605 / cat. 769 anti-AFP antibodies)
- Reference protein for the development of hepatocellular-carcinoma (HCC) biomarker assays, including AFP-L3 lectin-affinity workflows
- Reference protein for the development and calibration of prenatal-screening assays (maternal-serum AFP / MSAFP, amniotic-fluid AFP)
- Reference protein for the development and calibration of germ-cell-tumour (yolk-sac / endodermal-sinus tumour) assays
- Target antigen for bead-based combinatorial peptide-library screening and aptamer / scFv discovery (published — Jee 2014)
- Western-blot positive control and migration marker for AFP (~70 kDa)
- Ligand-binding studies — AFP binds fatty acids, bilirubin, retinoids, copper, and oestrogens; cord-blood native preparation retains the ligand-binding architecture
- Lens culinaris agglutinin (LCA) fractionation studies for AFP-L1 / AFP-L2 / AFP-L3 isoform separation
Cited use cases
The following peer-reviewed publication uses YO Proteins human alpha-Fetoprotein (cat. 858) as the target antigen of record.
1. Jee JE, Ang YL, Cha J, Ang MW, Ling J, Lim J, Lee SS. Combinatorial bead-based peptide libraries improved for rapid and robust screenings. Combinatorial Chemistry & High Throughput Screening. 2014;17(6):520–530. PMID 24410000.
Used YO Proteins human AFP (cat. 858) as the target antigen for an improved one-bead-one-compound (OBOC) combinatorial peptide-library screen. Demonstrates cat. 858 as a screening antigen for the discovery of synthetic peptide binders against a clinically important tumour-marker protein, validating its use in HCC-biomarker aptamer / peptide-mimic / capture-reagent development.
