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|Sample type||Average %||Range|
|Serum||99||92% - 103%|
|Cell culture media||118||116% - 119%|
|Heparin Plasma||103||102% - 105%|
|Citrate Plasma||94||86% - 100%|
TIMP2 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TIMP2 protein in mouse 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.
TIMP2 (Tissue inhibitor of metalloproteinases 2, Metalloproteinase inhibitor 2) is a widely expressed inhibitor of matrix metalloproteinases (MMPs). It functions by binding tightly to MMPs (in a 1:1 stoichiometry) and inactivating them by binding to their catalytic zinc cofactor. It is a soluble factor found circulating in serum and plasma. TIMP2 appears also have functions that are independent of MMP inhibition, including involvement in EGF signaling pathways.
|Components||1 x 96 tests|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|50X Cell Extraction Enhancer Solution (ab193971)||1 x 1ml|
|5X Cell Extraction Buffer PTR (ab193970)||1 x 10ml|
|Antibody Diluent BAR||1 x 6ml|
|Mouse TIMP2 Capture Antibody 10X||1 x 600µl|
|Mouse TIMP2 Lyophilized Recombinant||2 vials|
|Mouse TIMP2 Detector Antibody 10X||1 x 600µl|
|Plate Seals||1 unit|
|Sample Diluent LS||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 ab227893 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 TIMP2 were measured in duplicates, interpolated from the TIMP2 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:200, plasma (heparin) 1:200, and plasma (citrate) 1:200. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TIMP2 concentration was determined to be 519 ng/mL in serum, 548 ng/mL in plasma (heparin) and 448 ng/mL in plasma (citrate).
The concentrations of TIMP2 were measured in duplicates, interpolated from the TIMP2 standard curves and corrected for sample dilution. Undiluted samples are as follows: spleenocyte cell culture supernatant 25% and mouse lung cell culture supernatant 3%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TIMP2 concentration was determined to be 9.8 ng/mL in spleenocyte cell culture supernatant and 78.9 ng/mL in lung supernatant.
ab227893 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"