FACE™ Bad Chemi

by Active Motif
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Description

Fast Activated Cell-based ELISA (FACE™) Kits provide a simple, sensitive method for detecting protein phosphorylation directly in the cell, without making extracts or performing electrophoresis and membrane blotting. These 96-well, high-throughput assays are available in both colorimetric and chemiluminescent formats for over 20 different targets (see list at right). For complete details, click the FACE™ Method tab below.

FACE Bad Kits provide 96 rxns each of 2 antibodies that enable you to monitor and compare the levels of both phosphorylated and total Bad. The phospho-Bad antibody recognizes Bad only when phosphorylated at Ser112; the total-Bad antibody recognizes Bad regardless of the phosphorylation state. Click the Bad Info tab below for data and more information.

 
Name Format Cat No. Price  
FACE™ Bad 1 x 96 rxns 48165 $525 Buy Now
FACE™ Bad 5 x 96 rxns 48665 $2,100 Buy Now
FACE™ Bad Chemi 1 x 96 rxns 48265 $580 Buy Now
FACE™ Bad Chemi 5 x 96 rxns 48765 $2,320 Buy Now
Measurement of phosphorylated Bad and total Bad using the FACE Bad Cell Based ELISA (In Cell Western) method.
 
Figure 1: Measurement of phosphorylated and total Bad.

COS-7 cells were cultured in 96-well plates and serum-starved for 24 hours. Cells were then treated with 200 nM of PMA for 30 minutes and fixed. Total and phospho Bad were each assayed in triplicate using the phospho and total Bad antibodies included in the FACE Bad Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet).

Antibody Specificities

The phospho-Bad antibody is specific for phosphorylated Bad and was raised against a synthetic phospho-peptide corresponding to residues surrounding Serine 112 of mouse Bad. This antibody recognizes Bad only when phosphorylated at this site and does not cross-react with other sites or related family members. The total-Bad antibody recognizes Bad proteins regardless of the phosphorylation state.

Bad Overview

Bad (Bcl-2 antagonist of cell death) is a proapoptotic member of the Bcl-2 family that displaces Bax from binding to Bcl-2 and Bcl-xL, which results in cell death. Bad forms heterodimers with other Bcl-2 family members and localizes to the outer mitochondrial membrane. When phosphorylated in the presence of sur

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