孔雀石綠磷酸鹽測定試劑盒| Malachite Green Phosphate Assay Kit 貨號Abnova™ KA1331

Malachite Green Phosphate Assay Kit (Colorimetric, 600-660nm)-96-well plate、 384-well、 cuvette-based assays

孔雀石綠磷酸鹽測定試劑盒(Malachite Green Phosphate Assay Kit )提供了一種快速、可重複且非放射性(non-radioactive)的方法測量水溶液中的無機游離磷酸鹽(inorganic free phosphate)。這種簡單的測定方法是基於複雜的孔雀石綠鉬酸鹽( malachite green molybdate )與在酸性條件下形成游離正磷酸鹽(free orthophosphate)。孔雀石綠( malachite green-based assay)的測定方法，用於測量 free phosphate無機磷酸鹽水溶液。 該測定依賴於孔雀石綠之間形成複合物鉬酸鹽和free orthophosphate游離正磷酸鹽，溶液顏色從orange橙色變為綠色 green。顏色的形成可以方便地在分光光度計（620-640nm）上或在分光光度計上測量，讀值與游離正磷酸鹽濃度直接相關。不具放射性。該測定的應用包括磷酸化的定量以及從蛋白磷酸酶底物中釋放磷酸鹽。該測定僅測量不含無機物的磷酸鹽；脂質結合或蛋白質結合的磷酸鹽必須首先被水解和中和，然後才能測量。總體而言，該測定是檢測和量化最小限度的可靠且合適的方法。在酸性環境中含有大量的無機游離磷酸鹽，並且適合高通量篩選。

Figure 2. Typical standard curve in an Assay Buffer
Note: The Phosphate Standards #S1-S6 were diluted in Assay Buffer (50 mM HEPES, 1 mM DTT, 1 mM EDTA,
0.05% NP-40 with 0.1 mg/mL BSA) and run in triplicate (50 µL/well).

Figure 3. Typical standard curve in Milli-Q water
Note: The Phosphate Standards #S1-S6 were diluted in fresh Milli-Q water and run in triplicate (50 µL/well).

Table 2. Common Detergents and Reagents and their effect in the Malachite Green Assay.

Reagents were tested in the presence of 0, 0.08, 0.16, 0.31, 0.63, 1.25, and 2.5 nmol phosphate and compared to the Standard Curve diluted in UltraPure MilliQ water. Assay volumes were 50 µL prior to the addition MG Acidic and MG Blue solutions.

Publication Reference

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2.UPR proteins IRE1 and PERK switch BiP from chaperone to ER stress sensor.
Kopp MC, Larburu N, Durairaj V, Adams CJ, Ali MMU.Nat Struct Mol Biol. 2019 Nov;26(11):1053-1062. doi: 10.1038/s41594-019-0324-9. Epub 2019 Nov 6.

3.Transcriptional, translational, and physiological signatures of undernourished honey bees (Apis mellifera) suggest a role for hormonal factors in hypopharyngeal gland degradation.
Corby-Harris V, Meador CAD, Snyder LA, Schwan MR, Maes P, Jones BM, Walton A, Anderson KE.Journal of Insect Physiology. 2015 Dec 2.

4.SH2-PLA: a sensitive in-solution approach for quantification of modular domain binding by proximity ligation and real-time PCR.
Thompson CM, Bloom LR, Ogiue-Ikeda M, Machida K.BMC Biotechnol. 2015 Jun 26;15(1):60.