Abcam’s STAT3 in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of STAT3 protein in Human cells.
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.
|Components||1 x 96 tests|
|STAT3 (Total) Capture Antibody||1 x 3ml|
|STAT3 (Total) Detector Antibody||1 x 3ml|
|10X Wash Buffer PT||1 x 15ml|
|50X Cell Extraction Enhancer Solution||1 x 1ml|
|5X Cell Extraction Buffer PTR (ab193970)||1 x 10ml|
|Lyophilized STAT3 Control Lysate||1 vial|
|Plate Seal||1 unit|
|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 ab176655 in the following tested applications.
|Sandwich ELISA||Use at an assay dependent concentration.|
Example of a typical STAT3 cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Induction of STAT3 (pY705) phosphorylation in A431 cells in response to EGF treatment. A431 cells were cultured in 96-well tissue culture plates and treated (15 min) with a dose-range of EGF before cell lysis. Data from quadruplicate measurements of STAT3 (pY705) are plotted and compared against total STAT3 protein levels. Comparative STAT3 (pY705) and STAT3 (Total) data also shown by Western Blot.
Cell line analysis for Total STAT3 from 200 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1 x Cell Extraction Buffer PTR. Data from duplicate measurements of STAT3 (Total) are normalized and plotted.
ab176655 has not yet been referenced specifically in any publications.