Detects and quantifies the level of CREB (cAMP-Response Element-Binding protein) phosphorylated at Ser133 in human and mouse cells. CREB is a member of the leucine zipper family of DNA binding proteins. It induces transcription of target genes in response to several stimuli such as peptide hormones and growth factors.
| Product information | |||
|---|---|---|---|
| Format | 96-well plate | ||
| Form | 96 Tests | ||
| Detection method | Colorimetric | ||
| Species reactivity | human, mouse | ||
| Sensitivity | < 0.9 Units/ml | ||
| Assay range | 1.6-100 Units/ml | ||
| Assay time | 4 h | ||
| Kit contains | CREB (pSer¹³³) Standard, Standard Diluent Buffer, CREB Antibody-Coated 96-Well Plate, Rabbit Anti-CREB (pSer¹³³) Detector Antibody, Anti-Rabbit IgG-HRP Concentrate, HRP Diluent, Wash Buffer Concentrate, TMB, Stop Solution, Plate Covers and a user protocol. | ||
| Store and ship information | |||
|---|---|---|---|
| Storage | +2°C to +8°C | ||
| Ship |
Blue or Dry Ice
Multiple Toxicity Values, refer to MSDS |
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| Canadian export regulations | Due to the country and/or U.S. state of origin of the animal material used in this product, this product may not be exported to Canada. | ||
| Data | |||
|---|---|---|---|
   The sensitivity of this ELISA was compared to immunoblotting using known quantities of CREB (pSer<sup>133</sup>). The data show that the sensitivity of the ELISA is approximately 4x greater than that of immunoblotting. The bands shown in the immunoblot data were developed using rabbit anti- CREB (pSer<sup>133</sup>) and an alkaline phosphatase conjugated anti-rabbit IgG followed by chemiluminescent detection.  Natural CREB (pSer<sup>133</sup>) from Forskolin-treated HeLa cells was serially diluted in Standard Diluent Buffer. The absorbance of each dilution was plotted against the CREB (pSer<sup>133</sup>) standard curve. Parallelism was demonstrated by the figure above and indicated that the standard accurately reflects CREB (pSer<sup>133</sup>) content in samples.  HeLa cells were treated with 200 µM Forskolin for 20 min at 37°C. Untreated HeLa cells were used as control. All lysates were prepared and analyzed using the CREB (pSer<sup>133</sup>) ELISA Kit and CREB ELISA Kit. The total CREB remained comparable while the level of phosphorylated CREB increased after Forskolin treatment.  The specificity of this assay for phosphorylated CREB (pSer<sup>133</sup>) was confirmed by peptide competition. The data show that the phosphopeptide containing the phosphorylated Ser<sup>133</sup> blocks the ELISA signal. Non-phosphorylated peptide does not block the signal. |
