Description

Phosphatase Inhibitor Cocktail (100X) is a broad-spectrum phosphatase inhibitor mix designed to preserve protein phosphorylation status during cell lysis and protein extraction workflows. During sample preparation, endogenous phosphatases released from disrupted cells or tissues can rapidly remove phosphate groups from target proteins, resulting in reduced phosphoprotein signals and inaccurate analysis of phosphorylation-dependent signaling pathways. This 100X phosphatase inhibitor cocktail helps prevent protein dephosphorylation and maintains native phosphorylation states for downstream applications.

Formulated with commonly used phosphatase inhibitors including sodium fluoride, sodium pyrophosphate, β-glycerophosphate, and sodium orthovanadate, this protein phosphatase inhibitor cocktail provides broad-spectrum inhibition against endogenous serine/threonine phosphatases and protein tyrosine phosphatases. It is suitable for protecting phosphorylated proteins during preparation of cell lysates, tissue extracts, and other biological samples.

Phosphatase Inhibitor Cocktail (100X) is supplied as a convenient concentrated solution and is typically added to lysis buffers at a 1:100 dilution before sample extraction. Combined with protease inhibitors, it provides enhanced protection of protein samples for Western blot, phospho-Western blot, immunoprecipitation (IP), co-immunoprecipitation (Co-IP), and phosphorylation signaling pathway analysis.

Specifications

Product Name	Phosphatase Inhibitor Cocktail (100X)
SKU	AP0431
Volume	1 mL
Product Type	Phosphatase Inhibitor Cocktail
Concentration	100X
Working Dilution	1:100
Target Enzymes	Serine/Threonine Phosphatases, Tyrosine Phosphatases, Acid Phosphatases, Alkaline Phosphatases
Applications	Protein Extraction, Western Blot, Phospho-Western Blot, Immunoprecipitation, Co-IP
Sample Types	Cells, tissues, bacteria, fungi, plants
Storage	-20 °C
Shelf Life	12 months

Features

Broad-spectrum phosphatase inhibition: Effectively inhibits multiple endogenous phosphatases including serine/threonine and protein tyrosine phosphatases.
Preserves protein phosphorylation: Helps maintain phosphorylation states during cell lysis and protein extraction for accurate phosphoprotein analysis.
Convenient 100X concentrated formulation: Easy addition to RIPA buffer, NP-40 lysis buffer, and other protein extraction reagents at a 1:100 dilution.
Supports phospho-signaling research: Suitable for studies involving MAPK, PI3K/Akt, JAK/STAT, and other phosphorylation-dependent pathways.
Compatible with common protein workflows: Designed for Western blot, phospho-Western blot, immunoprecipitation, and protein interaction studies.

Applications

Phosphatase Inhibitor Cocktail (100X) is widely used as a protein extraction additive to protect phosphorylation-dependent protein modifications in molecular biology, cell biology, and biochemical research workflows.

Phospho-Western Blot Analysis: Preserves phosphorylated proteins during sample preparation to improve detection of phosphorylation-specific targets such as phospho-AKT, phospho-ERK, phospho-mTOR, and phospho-STAT proteins.
Cell Protein Extraction: Added to cell lysis buffers to inhibit endogenous phosphatase activity after cell disruption and maintain protein phosphorylation status.
Tissue Protein Extraction: Supports preparation of tissue lysates where endogenous phosphatases may affect phosphorylation analysis.
RIPA Lysis Buffer Supplement: Commonly combined with RIPA buffer and protease inhibitors for whole-cell protein extraction.
Immunoprecipitation (IP): Helps preserve phosphorylated epitopes during antibody-based protein enrichment workflows.
Co-Immunoprecipitation (Co-IP): Supports studies of phosphorylation-regulated protein interactions and signaling complexes.
Signal Transduction Research: Suitable for analysis of cellular signaling pathways regulated by reversible phosphorylation.
Protein Sample Preparation: Used for preparing stable protein lysates for biochemical analysis and downstream applications.

Protocol Overview

For optimal phosphoprotein preservation, Phosphatase Inhibitor Cocktail (100X) should be added to freshly prepared lysis buffer immediately before sample processing.

Add phosphatase inhibitor cocktail to lysis buffer at a 1:100 ratio to obtain the recommended working concentration.
For example, add 10 μL of 100X phosphatase inhibitor cocktail to 1 mL of lysis buffer.
Prepare inhibitor-containing lysis buffer immediately before use and avoid long-term storage after dilution.
Combine with protease inhibitor cocktail when protection against both protein degradation and dephosphorylation is required.
Perform cell or tissue lysis under cold conditions to minimize endogenous enzyme activity.

Ordering Information

SKU	Product	Size
AP0431	Phosphatase Inhibitor Cocktail (100X)	1 mL

FAQ

What is Phosphatase Inhibitor Cocktail (100X) used for?

Phosphatase Inhibitor Cocktail (100X) is used to inhibit endogenous phosphatases during protein extraction and cell lysis. It helps preserve protein phosphorylation states for Western blot, phospho-Western blot, immunoprecipitation, and signaling pathway analysis.

Why should phosphatase inhibitors be added to lysis buffer?

Cell lysis releases endogenous phosphatases that can remove phosphate groups from proteins and reduce phosphoprotein signals. Adding a phosphatase inhibitor cocktail helps prevent dephosphorylation and maintains the original phosphorylation status of target proteins.

How do you use a 100X phosphatase inhibitor cocktail?

The recommended dilution is 1:100. For example, add 10 μL of Phosphatase Inhibitor Cocktail (100X) to 1 mL of lysis buffer immediately before protein extraction.

Which phosphatases are inhibited by this cocktail?

This phosphatase inhibitor mix provides broad-spectrum inhibition against acid phosphatases, alkaline phosphatases, serine/threonine phosphatases, and protein tyrosine phosphatases.

Can this phosphatase inhibitor cocktail be used with RIPA buffer?

Yes. Phosphatase Inhibitor Cocktail (100X) can be added to RIPA lysis buffer before protein extraction. It is commonly used together with protease inhibitors for complete protection of protein samples.

What is the difference between protease inhibitor and phosphatase inhibitor?

Protease inhibitors prevent protein degradation caused by proteases, while phosphatase inhibitors prevent removal of phosphate groups from proteins. For phosphoprotein analysis, both inhibitors are often used together during cell lysis.

Which downstream applications are compatible with this product?

This product is suitable for Western blot, phospho-Western blot, SDS-PAGE sample preparation, immunoprecipitation, Co-IP, and phosphorylation signaling research workflows.