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HOME > Cell Biology > Cell signaling
MESG Phosphate Assay Kit
   
코    드 : 21659
단     위 : 200 assay
공 급 원 : AAT Bioquest
가     격 : 문의
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  MESG Phosphate Assay
PhosphoWorks™ Colorimetric MESG Phosphate Assay Kit
* UV absorption *


In the presence of inorganic phosphate MESG is converted to 2-amino-6-mercapto-7-methlpurine by purine nucleoside phosphorylase (EC 2.4.2.1) with absorption wavelength shift to red. This feature has been used to develop our convenient MESG phosphate assay kit. Our kit provides all the essential reagents including MESG, phosphorylase and reaction buffer. The MESG substrate gives an absorbance increase at 360 nm on phosphorylysis at pH 6.5-8.5, and at pH 7.6 the change in extinction coefficient is 11,000 M-1cm-1. The assay is shown to quantitate phosphate in solution at concentrations at least down to 2 µM. It can be used to measure the kinetics of phosphate release from phosphatases (such as GTPases and ATPases) by coupling the two enzymatic reactions


Key Features

Universal : Can be used for monitoring any biological processes that either generate or consume phosphate.
Continuous : Easily adapted to automation with no mixing or separation protocols.
Convenient : Formulated to have minimal hands-on time.
Non-Radioactive : No special requirements for waste treatment.
Use of Native substrates : Substrates can be proteins, peptides, nucleotides, sugars, organic molecules or inorganic salts.

Kit Components

Component A : Assay buffer                                        1 Bottle (10 mL)
Component B : MESG substrate                                   1 Vial (lyophilized powder)
Component C : Purine nucleosidephosphorylase (PNP) 1 Vial (lyophilized powder)
Component D : 1 mM KH2PO4                                      1 Vial (1 ml)

Brief Summary

→ Prepare 50 μL of test samples and/or phosphate standards
→ Add Assay Solution (50 μL)
→ Incubate at room temperature for 30 min
→ Read Absorbance at 360 nm

References

1. Webb MR, Hunter JL. (1992) Interaction of GTPase-activating protein with p21ras, measured using a continuous
    assay for inorganic phosphate release. Biochem J, 287 ( Pt 2), 555.
2. Webb MR. (1992) A continuous spectrophotometric assay for inorganic phosphate and for measuring phosphate
    release kinetics in biological systems. Proc Natl Acad Sci U S A, 89, 4884.
   
 
  MESG phosphate

Product Code

Unit

Price

Availability

21659

200 assay