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|Sample type||Average %||Range|
|Cell culture media||90||89% - 91%|
|Fetal Bovine Serum||57||48% - 61%|
|Bovine Serum Albumin||106||103% - 111%|
Abcam’s ZAP70 Human in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of catalase protein in Human 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.
ZAP70 is a tyrosine kinase that plays an essential role in the regulation of the adaptive immune response. ZAP70 regulates motility, adhesion and cytokine expression of mature T-cells, as well as in thymocyte development. ZAP70 regulates T-cell activation by modulating T-Cell Receptor expression at the cell surface.
|Components||1 x 96 tests|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|10X ZAP70 Capture Antibody||1 x 600µl|
|10X ZAP70 Detector Antibody||1 x 600µl|
|50X Cell Extraction Enhancer Solution (ab193971)||1 x 1ml|
|5X Cell Extraction Buffer PTR (ab193970)||1 x 10ml|
|Antibody Diluent CPI - HAMA Blocker (ab193969)||1 x 6ml|
|Plate Seals||1 unit|
|Sample Diluent NS||1 x 12ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
|ZAP70 Human Lyophilized Recombinant Protein||2 x 50ng|
Our Abpromise guarantee covers the use of ab173192 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.|
Example ZAP70 standard curve. Background-subtracted data values (mean +/- SD) are graphed. Data provided for demonstration purposes only. A new standard curve must be generated for each assay performed.
Titration of Jurkat extract within the working range of the Zap70 assay. Background subtracted data from duplicate measurements are plotted.
Interpolated values of ZAP70 are graphed for the indicated cells lines based on a extract load of 12.5 µg/mL. The ZAP70 detector antibody was used to analyze the same extracts by western blot (20 µg loaded/lane). The actin blot is included to show the relative loads of each lysate.
ab173192 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"