This Akt1 (phospho Thr72) Cell Based ELISA Kit allows for the detection of Akt1 (phospho Thr72) and the effects that certain stimulation conditions have on Akt1 (phospho Thr72) expression in different cell lines. Qualitative determination of Akt1 (phospho Thr72) concentration is achieved by an indirect ELISA format. In essence, the Akt1 (phospho Thr72) is captured by Anti-Akt1 (phospho Thr72) 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 Akt1 (phospho Thr72) 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-Akt1 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.
Piattaforma
Microplate
Tipo di rilevamento
Colorimetric
Tipo di campione
Adherent cells and suspension cells.
Campo di rilevamento
> 5000 Cells
Tempo del kit
4 hours 30 minutes
Reattività
Human, Mouse, Rat
Precision
Intra-assay CV: < 8%, Inter-assay CV: < 10%
Conservazione
Store at + 4°C. Stable for 6 months.
Sinonimi
AKT, AKT 1, AKT1_HUMAN, MGC99656, PKB, PKB-ALPHA, PRKBA, Protein kinase B, Protein Kinase B Alpha, Proto-oncogene c-Akt, RAC, RAC Alpha, RAC-alpha serine/threonine-protein kinase, RAC-PK-alpha
Dichiarazione di non responsabilità
Questo prodotto è solo per uso di ricerca. Non è inteso per uso diagnostico o terapeutico.
Figure 4: Western Blot - Akt1 (phospho Thr72) Cell Based ELISA Kit (CBP1240)
The mouse monoclonal antibody to GAPDH is used as a positive control inAkt1 (phospho Thr72) 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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