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HOME > Cell Biology > Cell signaling
Phosphate Assay Kit (Red)
   
코    드 : 21660
단     위 : 125 assay
공 급 원 : AAT Bioquest
가     격 : 문의
다운로드

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  Phosphate Assay kit
PhosphoWorks™ Fluorimetric Phosphate Assay Kit  *Red Fluorescence*


Cells utilize a wide variety of phosphate (Pi) and polyphosphate esters as enzyme substrates, second messengers, membrane structural components and vital energy reservoirs. Phosphate is involved in many biological processes. For example, phosphatases, ATPases and several other enzymes catalyze biochemical reactions in which inorganic phosphate is released from a phosphoester substrate. Detection of many phosphoester–metabolizing enzymes is difficult because suitable substrates are not available. It usually has been necessary to determine inorganic phosphate release using tedious colorimetric assays or radioisotope-based methods. This PhosphoWorks™ Fluorimetric Phosphate Assay Kit has been developed for measuring the activity of any Pi-generating enzyme using our red fluorescent phosphate sensor. The kit provides sensitive detection of Pi, an alternative to hazardous radioactive methods and other less sensitive colorimetric assays. The measurement of Pi is based on the change in the absorbance and fluorescence of our new phosphate sensor. Our kit provides all the essential reagents including phosphate sensor, phosphate standards and assay buffer. The assay is shown to quantitate phosphate in solution at concentrations at least down to 0.1 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. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation with no separation steps required.


Key Features

Dual Reading : Can be monitored by either fluorescence (Ex =540 nm / Em = 590 nm) or absorption (570 nm).
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.
Use of Native substrates : Substrates can be proteins, peptides, nucleotides, sugars, organic molecules or inorganic
                                     salts.

Kit Components

Component A : Assay buffer 1 Bottle (5 mL)                            -20 °C
Component B : Phosphate Sensor 1 Vial (lyophilized powder)   -20 °C and avoid light
Component C : 1 mM KH2PO4 1 Vial (100 ul)                            -20 °C

Brief Summary

→ Prepare test samples (40 μL) along with phosphate standard dilutions (40 μL) from Component C
→ Add equal volume of Component A (40 μL))
→ Add reconstituted Component B (20 μL)
→ Incubate at room temperature for 15 min to 1 hr
→ Read fluorescence using Ex 540 nm/Em 590 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.
   
 
  Phosphate

Product Code

Unit

Price

Availability

21660

125 assay