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5870
Phosphatase Inhibitor Cocktail (100X)
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Phosphatase Inhibitor Cocktail (100X) #5870

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Western Blotting Image 1: Phosphatase Inhibitor Cocktail (100X)

Western blot analysis of extracts from NIH/3T3 cells, prepared in lysis buffer in the absence of phosphatase inhibitors (left) or with Phosphatase Inhibitor Cocktail (100X) #5870 added (right), and incubated at 37ºC for the indicated time points, using Phospho-PKC (pan) (ΒII Ser660) Antibody #9371. In the absence of phosphatase inhibitors, Phospho-PKC signal fades within 3 hrs. after harvest, indicating protein degradation. In the presence of the phosphatase inhibitor cocktail, the Phospho-PKC degradation is slowed significantly and signal is still present at 24 hrs. following harvest.

Product Usage Information

Briefly vortex the Phosphatase Inhibitor Cocktail (100X) before use. Then just prior to use:

For Western Blot or Immunoprecipitation Cell Lysis: Dilute the cocktail 1:100 in desired lysis buffer to obtain a 1X working concentration.

For PTMScan(R) Cell Lysis: Dilute the cocktail 1:50 in urea lysis buffer to obtain a 2X working concentration. This higher concentration is required due to the concentrated nature of the lysate.

Solutions and Reagents

The Phosphatase Inhibitor Cocktail (100X) is composed of a proprietary mix of sodium fluoride, sodium pyrophosphate, β-glycerophosphate, and sodium orthovanadate to promote broad spectrum protection against endogenous serine/threonine and tyrosine phosphatases.

Storage

Store the undiluted 100X cocktail at 4ºC. Do not freeze.

Product Description

This Phosphatase Inhibitor Cocktail is used to prevent dephosphorylation of phosphorylated proteins from active serine/threonine and tyrosine phosphatases present in whole cell extract. The 100X cocktail is a colorless to light yellow liquid.

Background

Dynamic protein phosphorylation is a key cellular signaling mechanism by which a broad spectrum of cellular processes is regulated. In order to study the phosphorylation status of specific target proteins the phosphorylated residue of interest must remain intact. When cells are lysed to make whole cell extracts, a loss of normal cellular signaling regulation occurs, and phosphatases within the cell extract are free to dephosphorylate proteins in an uncontrolled manner. The addition of phosphatase inhibitors to the cell lysis buffer aids in the preservation of phosphorylated residues at the time of cell disruption.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PTMScan is a trademark of Cell Signaling Technology, Inc.
To Purchase # 5870S
Product # Size Price
5870S
1 ml $ 226