Smad3 (phospho Thr179) Cell Based ELISA Kit is a Enzyme-Linked Immunosorbent Assay designed for the indirect in vitro quantitative determination of Smad3 levels in adherent and suspension cells.
Assay Type
Cell-based (qualitative)
Principle of Assay
This Smad3 (phospho Thr179) Cell Based ELISA Kit allows for the detection of Smad3 (phospho Thr179) and the effects that certain stimulation conditions have on Smad3 (phospho Thr179) expression in different cell lines. Qualitative determination of Smad3 (phospho Thr179) concentration is achieved by an indirect ELISA format. In essence, the Smad3 (phospho Thr179) is captured by Anti-Smad3 (phospho Thr179) 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 Smad3 (phospho Thr179) 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-Smad3 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
Tissue Culture Treated Microplate
Detection Type
Colorimetric
Sample Type
Adherent and suspension cells.
Product Range
> 5000 Cells
Assay Time
4 hours 30 minutes
Reactivity
Human, Mouse, Rat
Storage
Store at + 4°C. Stable for 3-6 months.
Synonyms
hMAD-3, hSMAD3, JV15-2, MAD homolog 3, Mad3, MADH3, Mothers against decapentaplegic homolog 3, Mothers against DPP homolog 3, SMAD 3, SMAD family member 3
Product Note
Information online is representative. Always refer to the Product Manual inside the kit.
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
Western Blot - Smad3 (phospho Thr179) Cell Based ELISA Kit (A102187)
The mouse monoclonal antibody to GAPDH is used as a positive control inSmad3 (phospho Thr179) Cell Based ELISA Kit. Anti-GAPDH Antibody was tested for specificity by western blot with tissue lysates from human, mouse, and rat.
Proteins predicted to interact with Smad3
Predicted protein interactions based upon String database. Revelancy score correlates with probability of interaction.
