PhosphoBlocker Blocking Reagent

PhosphoBlocker Blocking Reagent

Cat. Num. AAA168232

• Synonyms: PhosphoBlocker Blocking Reagent; PhosphoBLOCKER Blocking Reagent; PhosphoBlocker Blocking Reagent reagent

• Concentration: 200 g dry blend; 5% concentration after reconstitution in 4L (varies by lot)

• Methods: Freshly prepare 5% PhosphoBLOCKER solution in TBST or PBST. Use the 5%; PhosphoBLOCKER solution to block the blot. When probing the blot, use the 5%; PhosphoBLOCKER solution to dilute primary and secondary antibodies.; Note: Reconstituted PhosphoBLOCKER solution is only good for one week at 4 degree C.
• Preparation and Storage: Room Temperature

Testing Data

Related Product Information for PhosphoBlocker Blocking Reagent reagent

• Superior blocking compared to dry milk or serum
• Enhances low level phosphoprotein signal without increasing background
• Preserves phosphoprotein antigens during Western blot procedures
• Easy-to-use premixed dry blend

Most commercially available Western blot blockers, such as dry milk or serum, are sufficient to block unreactive sites on the membrane. However, they are not designed to preserve phosphoprotein antigens during blotting. Our PhosphoBLOCKER Blocking Reagent provides superior blocking by maximizing signal-to-noise ratio. The PhosphoBLOCKER reagent particluarly excels with very low levels of endogenous phopsphoproteins.

Background: Protein phosphorylation-dephosphorylation is one of the major signaling mechanisms for modulating the functional properties of proteins involved in gene expression, cell adhesion, cell cycle, cell proliferation, and differentiation. Proteins can be phosphorylated by protein kinases on specific serine, threonine, or tyrosine residues. The utilization of anti-phosphoprotein antibodies in western blotting has become a commonly used tool for signal transduction research. Unfortunately, low levels of endogenous phosphoprotein in various cell lysates often can not be detected, even with high concentrations of antibody and long exposure times. Most commercially available western blot blockers (e.g. dry milk, serum) are sufficient to block the unreacted sites on the membrane, reducing the amount of nonspecific antibody binding during the assay; however, they are not designed to preserve phosphoprotein antigens during blotting.

MyBioSource's PhosphoBLOCKER contains a proprietary formulation that provides several advantages over conventional blockers:

Designed specifically for phosphoprotein blotting

Enhances low level phosphoprotein signal without increasing background

Premixed dry blend, easy to use

Product Categories/Family for PhosphoBlocker Blocking Reagent reagent

Cell Signaling and Protein Biology; Protein Analysis Tools; Protein Phosphorylation; PhosphoBLOCKER Blocking Reagent

References

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5. Ramakrishnan, R. et al. (2009). Characterization of Cdk9 T-loop Phosphorylation in Resting and Activated CD4+ T lymphocytes. J. Leukoc. Biol. 86:1345-1350.