FACE™ HSP27

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 HSP27 Kits provide 96 rxns each of 2 antibodies that enable you to monitor and compare the levels of both phosphorylated and total HSP27 (Heat Shock Protein 27). The phospho-HSP27 antibody recognizes HSP27 only when phosphorylated at Ser82. The total-HSP27 antibody recognizes HSP27 regardless of its phosphorylation state. Click the HSP27 Info tab below for data and more information.

 
Name Format Cat No. Price  
FACE™ HSP27 1 x 96 rxns 48350 $525 Buy Now
FACE™ HSP27 5 x 96 rxns 48850 $2,100 Buy Now
FACE™ HSP27 Chemi 1 x 96 rxns 48450 $580 Buy Now
FACE™ HSP27 Chemi 5 x 96 rxns 48950 $2,320 Buy Now
Measurement of phosphorylated HSP27 (S82) and total HSP27 using the FACE HSP27 Cell Based ELISA (In Cell Western) method.
 
Figure 1: Measurement of phosphorylated HSP27 (S82) and total HSP27.

Human HeLa cells were cultured in 96-well plates and serum-starved (0.5% FBS) for 16 hours. Cells were then stimulated with 25 µg/ml of Anisomycin for 30 minutes and fixed. Total HSP27 and HSP27 phosphorylated on Ser82 were each assayed in duplicate using the phospho-HSP27 (S82) and total-HSP27 antibodies included in the FACE HSP27 Kit. Data were plotted after correction for cell number (performed through use of Crystal Violet).

Antibody Specificities

The phospho-HSP27 (Ser82) antibody was raised against a synthetic phospho-peptide corresponding to residues surrounding phosphorylated Ser 82 of human HSP27. The phospho-HSP27 (Ser82) antibody recognizes HSP27 only when phosphorylated at Ser82. The total-HSP27 antibody recognizes HSP27 protein regardless of its phosphorylation state.

HSP27 Overview

In response to stress, the expression level of HSP27 increases several fold to confer cellular resistance to the adverse environmental change. HSP27 is localized to the cytoplasm of unstressed cells but can redistribute to the nucleus in response to stress, where it may function to stabilize DNA and/or the nuclear membrane. HSP27 is phosphorylated at serines 15, 78 and 82 by

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