|Sample type||Average %||Range|
|Cell culture media||101||101% - 102%|
|Extraction Buffer||103||100% - 107%|
PTP1B in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PTP1B protein in cell and tissue extracts.
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.
Protein tyrosine phosphatase, non-receptor type 1 (PTP1B) catalyse the hydrolysis of the phosphate monoesters specifically on tyrosine residues. PTP1B acts as a negative regulator of insulin signaling by dephosphylating the phosphotyrosine residues of insulin receptor kinase. PTP1B also dephosphoylate epidermal growth factor receptor kinase, JaK2, TYK2 kinases, PTP’s are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation.
|Components||1 x 96 tests|
|10X PTP1B Capture Antibody||1 x 600µl|
|10X PTP1B Detector Antibody||1 x 600µl|
|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 5BI||1 x 6ml|
|Plate Seals||1 unit|
|PTP1B Human Lyophilized Recombinant Protein||2 vials|
|Sample Diluent NS (ab193972)||1 x 12ml|
|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 ab184865 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.
Background subtracted data from duplicate measurements are plotted.
The concentrations of PTP1B protein were interpolated from the PTP1B standard curve and corrected for sample dilution relative to extract load of 333 µg/mL. The mean PTP1B concentration was determined to be 42.5 ng/mL in MCF7 extract at 333 µg/mL
Interpolated values of PTP1B are plotted for the indicated cell lines based on an extract load of 250 µg/mL. The PTP1B detector antibody was used to analyze the same extracts by western blot (20 µg loaded/lane).
ab184865 has not yet been referenced specifically in any publications.