This 4E-BP1 Cell Based ELISA Kit allows for the detection of 4E-BP1 and the effects that certain stimulation conditions have on 4E-BP1 expression in different cell lines. Qualitative determination of 4E-BP1 concentration is achieved by an indirect ELISA format. In essence, the 4E-BP1 is captured by Anti-4E-BP1 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 4E-BP1 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.
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
4EBP1, 4EBP1_HUMAN, BP 1, eIF4E binding protein 1, eIF4E-binding protein 1, eIF4EBP1, Eukaryotic translation initiation factor 4E-binding protein 1, PHAS-I, PHASI, Phosphorylated heat- and acid-stable protein regulated by insulin 1
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
Figure 4: Western Blot - 4E-BP1 Cell Based ELISA Kit (A102982)
The mouse monoclonal antibody to GAPDH is used as a positive control in4E-BP1 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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