This HDAC4 (phospho Ser632) Cell Based ELISA Kit allows for the detection of HDAC4 (phospho Ser632) and the effects that certain stimulation conditions have on HDAC4 (phospho Ser632) expression in different cell lines. Qualitative determination of HDAC4 (phospho Ser632) concentration is achieved by an indirect ELISA format. In essence, the HDAC4 (phospho Ser632) is captured by Anti-HDAC4 (phospho Ser632) Antibodies which in turn are detected by HRP-conjugated secondary antibodies. Through this binding, the HRP enzyme (conjugated to the secondary antibody) can catalyze a colorimetric reaction upon substrate addition. Due to the qualitative nature of this HDAC4 (phospho Ser632) Cell Based ELISA Kit, multiple normalization methods are described: 1) Anti-GAPDH Antibody is included to serve as an internal positive control in normalizing the target absorbance values. 2) Following the colorimetric measurement of HRP activity via substrate addition, the Crystal Violet whole-cell staining method is used to determine cell density. After staining, the results can be analyzed by normalizing the absorbance values to cell amounts, by which the plating difference can be adjusted. 3) Anti-HDAC4 Antibody is provided for normalization purposes. The absorbance values obtained for the non-phosphorylated target can be used to normalize the absorbance values for the phosphorylated target.
Platform
Microplate
Detection Type
Colorimetric
Sample Type
Adherent cells and suspension cells.
Product Range
> 5000 Cells
Assay Time
4 hours 30 minutes
Reactivity
Human, Mouse, Rat
Recovery
Storage
Store at + 4°C. Stable for 6 months.
Synonyms
AHO3, BDMR, EC 3.5.1.98, HA6116, HD 4, HD4, HDAC 4, HDAC A, HDAC4_HUMAN, HDACA, Histone deacetylase 4, Histone Deacetylase A, KIAA0288
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
Figure 1: Western Blot - HDAC4 (phospho Ser632) Cell Based ELISA Kit (A102204)
Western blot analysis of extracts from 293 cells treated with etoposide 25uM 1hour and Jurkat cells treated with etoposide 25uM 24hours, using Anti-HDAC4 (phopho Ser632) Antibody.
Figure 4: Western Blot - HDAC4 (phospho Ser632) Cell Based ELISA Kit (A102204)
The mouse monoclonal antibody to GAPDH is used as a positive control inHDAC4 (phospho Ser632) Cell Based ELISA Kit. Anti-GAPDH Antibody was tested for specificity by western blot with tissue lysates from human, mouse, and rat.
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