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CBA006  PhosphoDetect™ ERK1/2 (pThr¹85/pTyr¹87) ELISA Kit

p44 MAP Kinase, Phospho-Specific (Thr¹85/Tyr¹87) ELISA

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.
Product number Size Quantity Price
CBA006-1KIT  1 kit  price on request 
Alternatives
71296 PhosphoSafe™ Extraction Reagent 
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Alternative products for PhosphoDetect™ ERK1/2 (pThr¹85/pTyr¹87) ELISA Kit
71296 PhosphoSafe™ Extraction Reagent 
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Accessories for PhosphoDetect™ 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 or Dry Ice
Multiple Toxicity Values, refer to MSDS
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.

In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible).
R Phrase R: 21/22-36/38


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.

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