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SKU AR1183
Product nameBroad Spectrum Phosphatase Inhibitor Cocktail
Pack size 1 mL
Applications WB sample preparation, protein purification, Co-IP, protein kinase activity assay experiments

Overview

Form Supplied 100X concentrated stock solution
Physical State Liquid, colorless to light yellow
Pack Size 1 mL
Content Proprietary mix of: Sodium fluoride, Sodium orthavanadium, Imidazole, Sodium molybdate, Sodium sulphate, Sodium pyrophosphate, B-phosphoric acid glycerol
Recommended working concentration 100-fold dilution in tissue lysis buffer;
Use 10 µl of the Phosphatase Inhibitor Cocktail solution to inhibit dephosphorylation of proteins for 1 ml of lysate
Storage Upon receipt store at -20°C.
It is stable at -20°C for one year. It is stable at 4°C for two months. Product is shipped on ice.
Equivalent Thermofisher (Product No.78428, A32957); Millipore Sigma (Product No. P5726)
Target Specificity Tyrosine phosphatase, acidic and alkaline phosphatase; Serine/threonine phosphatase, histidine phosphatase, etc.

BioChemical Information

Inhibitor Inhibition Specificity
Sodium fluoride Ser/Thr and acid phosphatases
Sodium orthavanadium ATPases
Tyr and alkaline phosphatases
Imidazole Alkaline phosphatases
Sodium molybdate Acid and phosphoprotein phosphatases
Sodium tartrate Acid phosphatase
Sodium pyrophosphate Ser/Thr phosphatases
B-phosphoric acid glycerol Ser/Thr phosphatases

Assay Principle

Broad Spectrum Phosphatase Inhibitor Cocktail is a Western blot related concentrated stock solution reagent containing a mixture of different phosphatase inhibitors that is to be added to cell lysis buffer to protect native phosphoproteins from dephosphorylation during proteins purification and sample preparation used in WB, Co-IP, ChIP, and protein kinase assays.

Properties

Description Broad Spectrum Phosphatase Inhibitor Cocktail is a Western blot related concentrated stock solution reagent containing a mixture of six phosphatase inhibitors that is to be added to cell lysis buffer to protect native phosphoproteins from dephosphorylation during proteins purification and sample preparation used in WB, Co-IP, ChIP, and protein kinase assays.
Cite This Product Broad Spectrum Phosphatase Inhibitor Cocktail (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR1183)
Application WB sample preparation, protein purification, Co-IP, protein kinase activity assay experiments
Our Boster Guarantee covers the use of this product in the above tested applications.
Reagent Type Western Blotting related reagent; Inhibitors
Usage To preserve phosphorylation state and protein functionality following cell lysis
Target Sample Cell lysis extracts
Compatibility with assays Compatible with IEF/2D studies;
MS-compatible: not contain AEBSF;
Compatible with immobilized metal chelate affinity chromatography and 2D gel electrophoresis inhibitor components dialysis removal or desalting;

Usage and Handling

The product is supplied as a 100X concentrated stock solution in a liquid format for improved accuracy, solubility, and ease of use in comparison to traditional tablets. Since phosphatase levels may vary among cell and tissue types, it may be necessary to increase the concentration of inhibitors.

Background

Phosphatase inhibitors are used when phosphorylation (activation) states of target proteins need to be studied and the phosphorylated residues of interest must remain intact. They are chemicals that aid in the extraction of intact proteins in their native modification state by inhibiting endogenous phosphatases that would otherwise dephosphorylate the proteins present in cell lysates and tissue extracts. Broad Spectrum Phosphatase Inhibitor Cocktail contains individual components with specific inhibitory properties to provide an all-around protection of the protein phosphorylation state. The six phosphatase inhibitors included in this mixture target a broad spectrum phosphatase categories.
Dynamic protein phosphorylation is a key cellular signaling mechanism for cell processes regulation. When tissues are lysed to make whole cell extracts, the loss of natural compartmentalization causes normal regulation of cellular signaling to get distorted, and resident cell phosphatases within the cell extract are free to disorderly dephosphorylate proteins. The usual consequence of this unregulated state is biologically meaningless representation of protein activities (i.e. phosphorylation status) and false negative staining in anti-phosphoprotein immunostaining analyses. The addition of phosphatase inhibitors to the cell lysis buffer aids in the preservation of phosphorylated residues at the time of cell disruption.

Broad Spectrum Phosphatase Inhibitor Cocktail Images

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Broad Spectrum Phosphatase Inhibitor Cocktail
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