Non-Radioactive Phosphatase Assay Systems
Absorbance-Based Detection of Phosphatase Activity
- Measure serine/threonine phosphatase activity
- Use with purified samples, crude cell or tissue extracts
- Convenient alternative to radioactive phosphatase assays
Catalog Number:
Size
Catalog Number: V2460
Fast, Convenient Assays for Ser/Thr Phosphatase Activity
The Non-Radioactive Phosphatase Assay Systems determine the amount of free phosphate generated in a reaction by measuring the absorbance of a molybdate:malachite green:phosphate complex. They allow use of a variety of buffer conditions and substrates, including naturally phosphorylated proteins or synthetic phosphopeptides.
The effective range for the detection of phosphate released during an assay using the Phosphatase Assay Systems is 100–4,000pmol of phosphate.
Use with Purified Samples or Crude Cell and Tissue Extracts
In addition to measuring phosphatase activity in partially fractionated and purified samples, the Phosphatase Assay Systems can measure phosphatase activity in crude cell or tissue extracts. For this application, the high concentration of phosphate in these preparations is eliminated prior to performing the assay using the supplied Spin Columns, which rapidly and effectively remove free phosphate and other low-molecular-weight inhibitors from the sample. In addition, a unique Molybdate Dye Additive that is combined with the Molybdate Dye Solution aids in the solubilization of proteins exposed to the acid conditions of the Molybdate Dye Solution, which alone could potentially cause precipitation of the proteins.
Serine/Threonine Phosphatase Assay System
Contains the chemically synthesized phosphopetide, RRA(pT)VA, a peptide substrate that is compatible with several serine/threonine phosphatases such as the Protein Phosphatases 2A, 2B, and 2C. However the supplied phosphopeptide is a poor substrate for Protein Phosphatase 1 because of its more stringent structural requirements.
Protocols
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
|
Ser/Thr Phosphopeptide RRA(pT)VA |
V248A | 1 × 1mg |
|
Spin Column, Adaptor, and Reservoir |
V249A | 4 × 1 each |
|
Phosphate Standard |
V250A | 1 × 1ml |
|
96-Well Plate (1/2 area, flat bottom) |
V252A | 1 × 1 each |
|
Molybdate Dye Solution |
V253A | 1 × 20ml |
|
Molybdate Dye Additive |
V254A | 1 × 200μl |
|
Sephadex® G-25 |
V255A | 1 × 40ml |
|
Phosphate-Free Water |
V654A | 1 × 25ml |
SDS
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Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
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