|Sample type||Average %||Range|
|Serum||106||103% - 111%|
|Heparin Plasma||112||108% - 120%|
Abcam’s Insulin in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Insulin protein in Human serum and plasma (heparin).
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.
Insulin is a highly conserved, secreted hormone essential for glucose metabolism. Produced by pancreatic beta cells, proinsulin is proteolyzed into an A and a B chain, which form a 6 kDa mature protein. Basal levels of insulin are continuously delivered into the bloodstream, and additional levels are secreted proportional to food ingestion. Insulin secretion is highly regulated, and dysregulation of insulin production or sensitivity results in Type 1 diabetes mellitus or Type 2 diabetes mellitus, respectively.
|Components||1 x 96 tests|
|10X Human Insulin Capture Antibody||1 x 600µl|
|10X Human Insulin Detector Antibody||1 x 600µl|
|10X Wash Buffer PT||1 x 20ml|
|Antibody Diluent 5BI||1 x 6ml|
|Human Insulin Lyophilized Recombinant Protein||2 vials|
|Plate Seals||1 unit|
|Sample Diluent NS||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate||1 unit|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab200011 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Background-subtracted data values (mean +/- SD) are graphed.
Data shown for serum is from an individual donor with high endogenous insulin levels. Data shown for plasma is from a pooled (n=50) sample. Background-subtracted data values (mean +/- SD, n = 3) are graphed.
Ten individual healthy donors were evaluated for the presence of Insulin in serum using this assay. The range was from 11.8 – 345.1 pmol/L, with an average of 99.4 pmol/L. Health history and dietary status of donors were unknown.
ab200011 has not yet been referenced specifically in any publications.