|Sample type||Average %||Range|
|Cell culture supernatant||108||103% - 114%|
|Urine||94||89% - 99%|
|Serum||102||96% - 113%|
|Heparin Plasma||87||83% - 90%|
|EDTA Plasma||87||83% - 93%|
|Citrate Plasma||78||74% - 83%|
Abcam’s alpha Fetoprotein in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of alpha Fetoprotein protein in human serum, plasma and cell culture supernatant.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Samples diluted in Sample Diluent NS – 87 pg/mL
Samples diluted in Sample Diluent 25BS – 178 pg/mL
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.
|Components||1 x 96 tests|
|10X alpha Fetoprotein Capture Antibody||1 x 600µl|
|10X alpha Fetoprotein Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|alpha Fetoprotein Human Lyophilized Recombinant Protein||2 vials|
|Antibody Diluent 5BI||1 x 6ml|
|Plate Seals||1 unit|
|Sample Diluent 25BS||1 x 20ml|
|Sample Diluent NS (ab193972)||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab193765 in the following tested applications.
|Sandwich ELISA||Use at an assay dependent concentration.|
Background-subtracted data values (mean +/- SD) are graphed.
The Concentrations of alpha Fetoprotein were measured in duplicate and interpolated from the alpha Fetoprotein standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean alpha Fetoprotein concentration was determined to be 4.11ng/mL in normal human serum (n=50, 25 male and 25 female) and 219 ng/mL in pregnant human serum (n=1).
The concentrations of alpha Fetoprotein were measured in duplicate and interpolated from the alpha Fetoprotein standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean alpha Fetoprotein concentration was determined to be 219 ng/mL in pregnant human serum.
Serum from 10 apparently healthy male donors was measured in duplicate. The mean alpha Fetoprotein concentration was determined to be 2.57 ng/mL with a range of 0.54-7.58 ng/mL.