CBA006
PhosphoDetect™ ERK1/2 (pThr¹85/pTyr¹87) ELISA Kit
p44 MAP Kinase, Phospho-Specific (Thr¹85/Tyr¹87) ELISA
For general questions please contact our Customer Service:
EMD Chemicals, Inc.
480 S. Democrat Road
08027 Gibbstown
United States
Phone: 800 854 3417
Fax: 800 776 0999
E-mail: Customer.CareDesk@emdchemicals.com
11 February 2012
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Detects the dual phosphorylated forms of ERK1 at Thr202 and Tyr204 and ERK2 at Thr185 and Tyr187. This activation occurs as a result of treatment with a large variety of stimuli including mitogens, cytokines, and growth factors. Although this kit was designed for human samples, it cross-reacts with mouse and rat.
| Alternatives | |
|---|---|
| 71296 | PhosphoSafe™ Extraction Reagent |
| Alternative products for PhosphoDetect™ ERK1/2 (pThr¹85/pTyr¹87) ELISA Kit | |
| 71296 | PhosphoSafe™ Extraction Reagent |
| Accessories forPhosphoDetect™ ERK1/2 (pThr¹85/pTyr¹87) ELISA Kit |
| Product information | |||
|---|---|---|---|
| Format | 96-well plate | ||
| Form | 96 Tests | ||
| Detection method | Colorimetric | ||
| Species reactivity | human, mouse, rat | ||
| Avoid freeze/thaw | Yes | ||
| Sensitivity | ≤0.8 units/ml | ||
| Assay range | 1.6-100 units/ml | ||
| Assay time | 4 h | ||
| Sample type | Cell lysates | ||
| Kit contains | Coated 96-Well Plate, ERK1/2 Phospho-Thr¹85/Tyr¹87 Standard, Diluents, Detector Antibody, Secondary Antibody, Sample Treatment Buffer, Wash Buffer, TMB Substrate, Stop Solution, Plate Covers, and a user protocol. | ||
| Store and ship information | |||
|---|---|---|---|
| Storage | +2°C to +8°C | ||
| Ship |
Blue Ice Only
Multiple Toxicity Values, refer to MSDS |
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| Safety information | |||
|---|---|---|---|
| S Phrase |
S: 26-36/37-45 In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable protective clothing and gloves. In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible). |
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| R Phrase |
R: 21/22-36/38 Harmful in contact with skin and if swallowed. Irritating to eyes and skin. |
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| Data | |||
|---|---|---|---|
 Jurkat cells were treated with 100 ng/ml PMA for 10 min and cell lysates were prepared. Cell lysate prepared with Cell Lysis Buffer was either boiled for 5 min, treated with Sample Treatment Buffer, or not treated. Cell lysate was also prepared using Denaturing Cell Lysis Buffer. Samples were analyzed with the ERK1/2 and the PhosphoDetect™ ERK (Thr<Sup>185</sup>/Tyr<sup>187</sup>) ELISA Kits.  The sensitivity of this ELISA was compared to immunoblotting using known quantities of ERK1/2 Phospho-Thr<sup>185</sup>/Tyr<sup>187</sup>. The data shows that the sensitivity of the ELISA is approximately 4X greater than that of immunoblotting. The bands shown in the immunoblotting were developed using rabbit anti-ERK1/2 Phospho-Thr<sup>185</sup>/Tyr<sup>187</sup> and an alkaline phosphatase conjugated anti-rabbit IgG followed by chemiluminescent detection.  Natural ERK1/2 Phospho-Thr<sup>185</sup>/Tyr<sup>187</sup> from PMA-treated Jurkat cell lysates was serially diluted in Standard Diluent Buffer. The absorbance of each dilution was plotted against the ERK1/2 Phospho-Thr<sup>185</sup>/Tyr<sup>187</sup> standard curve. Parallelism is demonstrated in the figure above and indicates that the standard accurately reflects natural ERK1/2 Phospho-Thr<sup>185</sup>/Tyr<sup>187</sup> content in samples. |
