|Pack size||80mg (sufficient for 100mL of extract)|
|Physical State||White lyophilized powder|
|Applications||Animal tissue and cell extract with detergent|
|Product Name||Phosphatase Inhibitor Cocktail III|
|Physical State||White lyophilized powder|
|Pack Size||80mg (sufficient for 100mL of extract)|
|Content||Proprietary mix of: Sodium fluoride, Sodium orthavanadium, Sodium pyrophosphate, B-phosphoric acid glycerol|
|Recommended working concentration||Reconstitute Phosphatase Inhibitor Cocktail III with 1mL of water.|
|Storage||Upon receipt store at -20°C. It is stable for one year. Product is shipped on ice.|
|Equivalent||Thermofisher (Product No. 78428, A32957)|
|Type of Phosphatase Inhibitor Cocktail||Specificity||Catalog Number|
|Phosphatase Inhibitor Cocktail II||Tyrosine protein phosphatases, acid and alkaline phosphatases||AR1120|
|Phosphatase Inhibitor Cocktail III||Serine phosphatases, threonine phosphatases and protein tyrosine phosphatases||AR1121|
|Sodium fluoride||Acid phosphatase, Serine phosphatases, Threonine phosphatases|
|Sodium orthavanadium||Alkaline phosphatases, Tyrosine phosphatase|
|Sodium pyrophosphate||Serine phosphatases, Threonine phosphatases|
|B-phosphoric acid glycerol||Serine phosphatases, Threonine phosphatases|
Crude cell or tissue extracts contain a number of endogenous enzymes, such as proteases and phosphatases, which are capable of modifying the proteins present in the extracts. Inhibitors of these enzymes should be added to improve the yield of native proteins. Phosphatase inhibitor cocktail III protect phosphoproteins from serine phosphatases, threonine phosphatases and protein tyrosine phosphatases. The cocktail contains a mixture of four inhibitors, including sodium fluoride, sodium orthovanadate, beta-sodium glycerophosphate and sodium pyrophosphate. It is suitable for use with many kinds of cell and tissue extract, including primary cells, mammalian cells, animal tissue, plant tissue, yeast or bacterial cells.
|Compatibility with reagents||Fully compatible with cell lysis buffers and Broad Spectrum Protease Inhibitor Cocktail|
|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;
|Reagent Type||Western Blotting related reagent; Inhibitors|
|Usage||To preserve phosphorylation state and protein functionality following cell lysis|
|Target Specificity||Serine phosphatases, threonine phosphatases and protein tyrosine phosphatases|
|Target Sample||Cell lysis extracts|
|Description||Boster’s Phosphatase Inhibitor Cocktail III is a Western blot related 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, and IP.|
|Cite This Product||Phosphatase Inhibitor Cocktail III (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR1121)|
|Application||Animal tissue and cell extract with detergent
*Our Boster Guarantee covers the use of this product in the above tested applications.
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. Phosphatase Inhibitor Cocktail III contains individual
components with specific inhibitory properties to provide an all-
around protection of the protein phosphorylation state.
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.
• Dialyzable—inhibitors can be removed by dialysis, before
performing assays known to be incompatible with phosphatase
• Supplied as a 100X concentrate—effective as 1X. For samples with high level of phosphatase may need 2-3X.
• Detergent compatible—can be used with lysis buffers and do not crossreact with detergents in the buffers.
• Protein Assay Compatible—works with BCA and commassie blue assays.
Usage and Handling
The product is supplied as white lyophilized powders. Since phosphatase levels may vary among cell and tissue types, it may be necessary to increase the concentration of inhibitors.
1. Reconstitute Phosphatase Inhibitor Cocktail III in 1ml of
double distilled water.
2. Add cocktail solution to extraction buffer in 1:100. (e.g., Add 10 μL of cocktail solution to 1mL extraction buffer.)
3. Proceed to extract protein fractions.
Notes:Aliquot and store reconstituted phosphatase inhibitor cocktail solution at -20˚C.
Phosphatase Inhibitor Cocktail III Images
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