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

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

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15 Februar 2012

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CBA006-1KIT  1 kit 
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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.
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71296 PhosphoSafe™ Extraction Reagent
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71296 PhosphoSafe™ Extraction Reagent
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Zubehör fürPhosphoDetect™ ERK1/2 (pThr¹85/pTyr¹87) ELISA Kit
Produktinformationen
Format 96-well plate
Form 96 Tests
Detection method Colorimetric
Species reactivity human, mouse, rat
Avoid freeze/thaw Ja
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.
Lager- und Versandinformationen
Lagerkategorie +2°C to +8°C
Ship Blue Ice Only
Multiple Toxicity Values, refer to MSDS
Sicherheitshinweise
S-Satz S: 26-36/37-45



R-Satz R: 21/22-36/38


Daten

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.

© Merck KGaA, Darmstadt, Germany, 2012

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