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|Sample type||Average %||Range|
|Serum||105||104% - 108%|
|Cell culture media||103||100% - 108%|
|Heparin Plasma||97||94% - 102%|
|EDTA Plasma||108||102% - 117%|
|Citrate Plasma||98||95% - 101%|
ab187394 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TIMP Metalloproteinases Inhibitor 1 (TIMP1) protein in human serum, plasma, and cell culture supernatant samples.
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.
Tissue inhibitor of metalloproteinases 1 (TIMP1 or Metalloproteinase inhibitor 1) is a widely expressed inhibitor of matrix metalloproteinases (MMPs). It functions by binding non-covalently to MMPs (in a 1:1 stoichiometry) and blocking access of substrate to the MMP active site. TIMP1 expression is induced by pro-inflammatory cytokines. It is a soluble factor found circulating in serum and plasma. TIMP1 appears also have functions that are independent of MMP inhibition, including promoting erythropoiesis and inhibiting apoptosis in B cells.
|Components||1 x 96 tests|
|10X TIMP-1 Capture Antibody||1 x 600µl|
|10X TIMP-1 Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|Antibody Diluent CPI - HAMA Blocker (ab193969)||1 x 6ml|
|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|
|TIMP-1 Human Lyophilized Recombinant Protein||2 vials|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab187394 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.|
The TIMP1 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.
PBMC were grown in the absence or presence of phytohemagglutinin (PHA) for 2 days. TIMP1 was measured in 2, 4, 8, 16 and 32-fold diluted cell culture supernatants of unstimulated and PHA stimulated PBMC. Raw values (Mean +/-SD, n=2) are graphed.
Background subtracted data from duplicate measurements are plotted.
Serum from 10 apparently healthy male donors was measured in triplicate. The mean TIMP1 concentration was determined to be 109 ng/mL with a range of 85 – 125 ng/mL.
PBMC were grown in the absence (unstimulated) or presence of phytohemagglutinin (PHA) (stimulated) for 2 days (stimulated). TIMP1 was measured in 2-fold diluted cell culture supernatants of unstimulated and PHA stimulated PBMC. Measured values were interpolated from the TIMP1 Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed: 10 ng/mL unstimulated and 8.2 ng/mL stimulated.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"