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|MCF-7 cells||< 10%|
p53 (TP53 gene) acts as a tumor suppressor in many tumor types and induces growth arrest or apoptosis depending on the physiological circumstances and cell type. p53 is involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. p53 mediated apoptosis induction seems to be by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. p53 is also implicated in Notch signaling cross-over.
In-Cell ELISA (ICE) technology is used to perform quantitative immunocytochemistry of cultured cells with a near-infrared fluorescent dye-labeled detector antibody. The technique generates quantitative data with specificity similar to Western blotting, but with much greater quantitative precision and higher throughput due to the greater dynamic range and linearity of direct fluorescence detection and the ability to run 96 samples in parallel. Because the p53 antibody is a mouse antibody and the p53 phospho S46 antibody is a rabbit antibody, their targets can be measured simultaneously in a same well using the cocktail of provided primary antibodies and the provided cocktail of IRDye®-labeled species-specific secondary antibodies when using a LI-COR infrared imager. This method rapidly fixes the cells in situ, stabilizing the in vivo levels of proteins and their post-translational modifications, and thus essentially eliminates changes during sample handling, such as preparation of protein extracts. Finally, the p53 phospho Ser46 and total p53 protein signals can all be normalized to cell amount, measured by the provided Janus Green whole cell stain, to further increase the assay precision.
Plates are available in our ICE (In-Cell ELISA) Support Pack (ab111542) which can be bought seperately.
|Components||1 x 96 tests|
|1000X IRDye® Labeled Secondary Antibody Cocktail||1 x 24µl|
|100X Mouse Anti-p53 Primary Antibody||1 x 120µl|
|100X Rabbit Anti-p53 Phospho Ser-46 Primary Antibody||1 x 120µl|
|100X Triton X-100||1 x 0.5ml|
|10X Blocking Buffer||1 x 10ml|
|10X Phosphate Buffered Saline||1 x 100ml|
|1X Janus Green Stain||1 x 11ml|
|400X Tween-20||1 x 2ml|
Our Abpromise guarantee covers the use of ab128570 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|In-Cell ELISA||Use at an assay dependent concentration.|
ab128570 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"