*This blog is created based on the frequent question about Phosphatase Inhibitor Cocktail III(P-1549) application from our customers.

Q. How much volume of stock solution will be added to a certain amount of cell lysate?

A. 1mL of the stock solution is good for 500 mg of protein extracted from the tissue or cells. Here is the Recommended Usage. Recommended for use with tissue and cell extracts including extracts containing detergents. Reconstitute each vial with 1 ml H2O to obtain a 1 ml stock solution

Q. How about 800uL lysis buffer to lysate 10x10 cm2 cell?

A. With an unkown amount of protein his is hard to be exact with a recommendation. It is essential to use too much then to use too little phosphatase inhibitor.

Description: When diluted in lysis buffer to a final concentration of 1X the 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.

Directions for Use:

1. Briefly vortex the Phosphatase Inhibitor Cocktail (100X) before use.

2. Just prior to lysing cells, dilute the cocktail 1:100 in desired lysis buffer to obtain a 1X working concentration

3. Solutions and Reagent: 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.

4. 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.

5. After reviewing some other papers use of 1 vial, prepared as stated above, inhibits> 96% of the phosphatases. We would offer this as researchers starting point.

 

Specific Inhibitors, Inhibitor Cocktail

 

At AG Scientific we use only the necessary ingredients helps eliminate contamination and lower your costs, ultimately resulting in more efficient and reproducible research.

 

Inhibitor Cocktail

Components

Specificity of Inhibition

Recommended Applications

Protease Inhibitor Cocktail I AEBSF, Aprotinin, E-64, EDTA, Leupeptin Serine, cysteine, and trypsin-like proteases; esterases; metalloproteases General Use
Protease Inhibitor Cocktail I, Animal-Free AEBSF, Aprotinin(recombinant), E-64, EDTA, Leupeptin Serine, cysteine, and trypsin-like proteases; esterases; metalloproteases General use and applications that require animal-free reagents
Protease Inhibitor Cocktail II AEBSF, bestatin, E-64, EDTA, pepstatin A Serine, cysteine, and aspartic proteases; aminopeptidases; metalloproteases Bacterial cell extracts
Protease Inhibitor Cocktail III  AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A Serine, cysteine, aspartic, and trypsin like proteases; aminopeptidases Mammalian cells and tissue extracts
Protease Inhibitor Cocktail III, Animal-Free  AEBSF, aprotinin(recombinant), bestatin, E-64, leupeptin, pepstatin A Serine, cysteine, aspartic, and trypsin-like proteases; aminopeptidases Mammalian cells and tissue extracts and applications that require animal-free reagents
Protease Inhibitor Cocktail III, Animal-Free, DMSO-Free AEBSF, aprotinin(recombinant), bestatin, E-64, leupeptin, pepstatin A Serine, cysteine, aspartic, and trypsin-like proteases; aminopeptidases Mammalian cells and tissue extracts and animal-free, organic solvent free applications
Protease Inhibitor Cocktail IV AEBSF, E-64, Pepstatin A, Phenanthroline Serine, cysteine, and aspartic proteases; metalloproteases Fungal and yeast cell extracts
Protease Inhibitor Cocktail V, EDTA-free  AEBSF, Aprotinin, E-64, Leupeptin Serine, cysteine, and trypsin-like proteases Mammalian cells and tissue extracts, samples analyzed by 2-D gel electrophoresis
Protease Inhibitor Cocktail V, EDTA-free, animal-free  AEBSF, Aprotinin(recombinant),E-64, Leupeptin Serine, cysteine, and trypsin-like proteases Mammalian cells and tissue extracts and applications that require animal-free reagents
Protease Inhibitor Cocktail VI, General Use  AEBSF, aprotinin, bestatin, E-64, EDTA, leupeptin Serine, cysteine, and trypsin-like protease; aminopeptidases; metalloproteases General Use
Protease Inhibitor Cocktail VI, Plant Cells  AEBSF, bestatin, E-64, leupeptin, phenanthroline, pepstatin A Serine, cysteine, and aspartic proteases; aminopeptidases; metalloproteases Plant cell extracts
Protease Inhibitor Cocktail VII AEBSF, bestatin, E-64, pepstatin A, phosphoramidon Serine, cysteine, and aspartic proteases; aminopeptidases; metalloendopeptidases Histidine-tagged proteins
Protease Inhibitor Cocktail VII, DMSO-free  AEBSF, bestatin, E-64, pepstatin A, phosphoramidon Serine, cysteine, and aspartic proteases; aminopeptidases; metalloendopeptidases Histidine-tagged proteins and organic solvent free applications
Protease Inhibitor Cocktail VIII  ALLN, antipain, E-64 Calpain I/II, cathepsin A/B, cathepsin L/S, papain, cysteine proteases Broad range cysteine protease inhibition
Serine Protease Inhibitor Cocktail I AEBSF, aprotinin, elastatinal, GGACK Cymotrypsin, kallikrein, plasmin, thrombin, trypsin, elastase, urokinase, Factor Xa Broad range serine protease inhibition
Phosphatase Inhibitor Cocktail I  Bromotetramisole, cantharidin, microcystin LR Alkaline phosphatases, protein phosphatase 1/2A(PP-1/PP-2A) Animal tissues, A431 or Jurkat cell extracts
Phosphatase Inhibitor Cocktail II Imidazole, Na(orthovanadate, fluoride, tartrate, molybdate) Protein tyrosine phosphatase, acid phosphatases, alkaline phosphatases Animal tissues, A431 or Jurkat cell extracts
Phosphatase Inhibitor III Na(orthovanadate, fluoride, pyrophosphate), glycerophosphate Ser/Thr phosphatases, acid phosphatases, alkaline phosphatases, PP-1/PP-2A Animal tissue extracts
Phosphatase Inhibitor Cocktail IV  Bromotetramisole, cantharidin, calyculin A Alkaline phosphatases, PP-1/PP-2A Animal tissue extracts
PIA 100x, Protease Inhibitor A Antipain, leupeptin, 50mg/ml EDTA in water Ca2+-dependent endopeptidases, papain, trypsin-like serine and cysteine proteases Cysteine protease and papain inhibition, custom concentration 1
PIA 1000x, Protease Inhibitor A Antipain, leupeptin, 50mg/ml EDTA in water Ca2+-dependent endopeptidases, papain, trypsin-like serine and cysteine poteases Cysteine protease and papain inhibition, custom concentration 2

 

[Get your Free Inhibitor Brochures here]

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10 Items

per page
  1. Protease Inhibitor Cocktail I | AEBSF,Aprotinin, E-64, EDTA,Leupeptin
    Protease Inhibitor Cocktail I
    P-1510

    Starting at $39.08

  2. Protease Inhibitor Cocktail II | Inhibitors | AG Scientific - proteomic biomarker preservation agent contains AEBSF.HCl Bestatin E-64 EDTA Pepstatin A
    Protease Inhibitor Cocktail II
    P-1511

    Starting at $53.61

  3. Protease Inhibitor Cocktail III | Inhibitors | AG Scientific
    Protease Inhibitor Cocktail III
    P-1512

    Starting at $50.26

  4. Protease Inhibitor Cocktail IV | Inhibitors | AG Scientific
    Protease Inhibitor Cocktail IV
    P-1513

    Starting at $60.72

  5. Protease Inhibitor Cocktail V, EDTA-free | Inhibitors | AG Scientific
    Protease Inhibitor Cocktail V, EDTA-Free
    P-1514

    Starting at $107.79

  6. Protease Inhibitor Cocktail VI (General Use) | AG Scientific
    Protease Inhibitor Cocktail VI
    P-1515

    Starting at $39.12

  7. Protease Inhibitor Cocktail VIII | Cysteine Protease | AG Scientific
    Protease Inhibitor Cocktail VIII
    P-1548

    Starting at $67.00

  8. Phosphatase Inhibitor Cocktail III | AG Scientific
    Phosphatase Inhibitor Cocktail III
    P-1549

    Starting at $44.69

  9. Serine Protease Inhibitor Cocktail I inhibits serine proteases contains AEBSF. HCl, 420 nM Aprotinin, 20 µM Elastatinal and 1µM GGACK.
    Serine Protease Inhibitor Cocktail I
    P-1516

    Starting at $95.55

  10. Protease Inhibitor Cocktail VI, DETA-Free | AG Scientific, Inc.
    Protease Inhibitor Cocktail VI, EDTA-Free
    P-1554

    Starting at $39.08

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10 Items

per page