Human alpha Fetoprotein ELISA Kit (ab193765)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 44.8 pg/ml
- Range: 156.25 pg/ml - 10000 pg/ml
- Sample type: Cell culture media, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
-
Product name
Human alpha Fetoprotein ELISA Kit
See all alpha 1 Fetoprotein kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 8 2% Inter-assay Sample n Mean SD CV% Serum 3 4.5% -
Sample type
Cell culture supernatant, Urine, Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
44.8 pg/ml -
Range
156.25 pg/ml - 10000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 103 101% - 113% Urine 119 116% - 121% Serum 91 88% - 96% Cell culture media 94 91% - 98% Hep Plasma 96 94% - 97% EDTA Plasma 100 98% - 102% Cit plasma 89 89% - 89% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human alpha Fetoprotein ELISA Kit (ab193765) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of alpha Fetoprotein protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, urine, and cell culture media. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human alpha Fetoprotein with 44.8 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
-
Notes
Alpha-fetoprotein (AFP) is a secreted single chain glycoprotein expressed normally during fetal development as well as by liver pathologies, including cancer. It is a member of the albuminoid superfamily and may act as a carrier protein. Pregnant women may have elevated alpha-fetoprotein levels derived from fetal circulation and male and female adults have low levels of circulating alpha-fetoprotein.
-
Platform
Microplate (12 x 8 well strips)
Properties
-
Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 5BI 1 x 6ml Human alpha Fetoprotein Capture Antibody (Lyophilized) 1 vial Human alpha Fetoprotein Detector Antibody (Lyophilized) 1 vial Human alpha Fetoprotein Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent 75BS 2 x 20ml Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
-
Function
Binds copper, nickel, and fatty acids as well as, and bilirubin less well than, serum albumin. Only a small percentage (less than 2%) of the human AFP shows estrogen-binding properties. -
Tissue specificity
Plasma. Synthesized by the fetal liver and yolk sac. -
Sequence similarities
Belongs to the ALB/AFP/VDB family.
Contains 3 albumin domains. -
Developmental stage
Occurs in the plasma of fetuses more than 4 weeks old, reaches the highest levels during the 12th-16th week of gestation, and drops to trace amounts after birth. The serum level in adults is usually less than 40 ng/ml. AFP occurs also at high levels in the plasma and ascitic fluid of adults with hepatoma. -
Post-translational
modificationsIndependent studies suggest heterogeneity of the N-terminal sequence of the mature protein and of the cleavage site of the signal sequence.
Sulfated. -
Cellular localization
Secreted. - Information by UniProt
-
Alternative names
- Afp
- AFPD
- Alpha fetoglobulin
see all -
Database links
- Entrez Gene: 174 Human
- Omim: 104150 Human
- SwissProt: P02771 Human
- Unigene: 518808 Human
Images
-
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
-
Standard Curve comparison between human AFP SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.
-
The AFP standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
-
The concentrations of AFP were measured in duplicates, interpolated from the AFP standard curves and corrected for sample dilution. Undiluted samples are as follows: pregnant human serum 3% and HepG2 cell culture supernatant 1:666. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration was determined to be 221.8 ng/mL in neat pregnant human serum and 6,165 ng/mL in neat HepG2 cell culture supernatant.
-
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration was determined to be 678.4 pg/mL with a range of 133.9 – 2,257 pg/mL.
-
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration in nine detectable donor samples was determined to be 550.7 pg/mL with a range of 247.9-816.1 pg/mL. Donor #5 was not detectable.
-
To learn more about the advantages of recombinant antibodies see here.