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|Sample type||Average %||Range|
|Serum||95||90% - 99%|
|Cell culture media||116||114% - 119%|
|Heparin Plasma||116||108% - 122%|
|EDTA Plasma||106||103% - 108%|
|Citrate Plasma||109||104% - 111%|
Abcam’s Leptin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of Leptin protein in Human serum, plasma and cell culture supernatants.
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.
Leptin is a secreted protein factor produced by adipocytes. It regulates energy balance and body fat deposits. Leptin deficiency in humans and mice can cause in obesity.Acute low levels of leptin triggers increased appetite and chronic leptin deficiency can block the onset of puberty.
|Components||1 x 96 tests|
|10X Leptin Capture Antibody||1 x 600µl|
|10X Leptin Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|Antibody Diluent CPI - HAMA Blocker (ab193969)||1 x 6ml|
|Leptin Human Lyophilized Recombinant||2 vials|
|Plate Seals||1 unit|
|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 ab179884 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.
The concentrations of Leptin were interpolated from the Leptin standard curve and corrected for sample dilution. The mean Leptin concentration was determined to be 6,260 pg/mL in plasma (citrate) and 1,845 pg/mL in serum.
Serum from 10 apparently healthy male donors was measured in triplicate for Leptin. The mean Leptin concentration was determined to be 2174 pg/mL with a range of 443 – 6250 pg/mL (Leptin was not detected in serum from Donor 7).