跳转至内容
Merck
CN

In Gel Kinase Assay.

Bio-protocol (2017-03-05)
Gastón A Pizzio, Pedro L Rodriguez
摘要

Proper spatiotemporal regulation of protein phosphorylation in cells and tissues is required for normal development and homeostasis. We present the protocol 'In Gel Kinase Assay', which is useful for protein kinase activity measurements from crude protein extracts. We have successfully used 'In Gel Kinase Assay' protocol to show that the Arabidopsis thaliana sextuple mutant in the PYRABACTIN RESISTANCE1/PYR1-LIKE/REGULATORY COMPONENTS OF ABA RECEPTORS (PYR/PYL/RCAR-ABA receptors; line pyr/pyl112458) is impaired in ABA-mediated activation of SnRK2.2, SnRK2.3 and OST1/SnRK2.6, as much as the triple mutant snrk2.2/2.3/2.6 (Gonzalez-Guzman et al., 2012).

材料
产品编号
品牌
产品描述

Sigma-Aldrich
甘油, Molecular Biology, ≥99.0%
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
十二烷基硫酸钠, BioReagent, suitable for electrophoresis, Molecular Biology, ≥98.5% (GC)
Sigma-Aldrich
MES 水合物, ≥99.5% (titration)
Sigma-Aldrich
TWEEN ® 20, viscous liquid
Sigma-Aldrich
Triton X-100, Molecular Biology
Sigma-Aldrich
乙二胺四乙酸 二钠盐 二水合物, suitable for electrophoresis, Molecular Biology, 99.0-101.0% (titration)
Sigma-Aldrich
氯化镁, ≥98%
Sigma-Aldrich
苯甲磺酰氟, ≥98.5% (GC)
Sigma-Aldrich
三氯乙酸, ACS reagent, ≥99.0%
Sigma-Aldrich
三(羟甲基)氨基甲烷, ACS reagent, ≥99.8%
Sigma-Aldrich
β-甘油磷酸盐 二钠盐 水合物, BioUltra, suitable for cell culture, suitable for plant cell culture, ≥99% (titration)
Sigma-Aldrich
SIGMAFAST 蛋白酶抑制剂混合物片剂,无EDTA, for use in purification of Histidine-tagged proteins
Sigma-Aldrich
乙二醇-双(2-氨基乙醚)-N,N,N′,N′-四乙酸, Molecular Biology, ≥97.0%
Sigma-Aldrich
腺苷 5′-三磷酸腺苷 (ATP) 二钠盐 水合物, vial of 30 mg
Sigma-Aldrich
正钒酸钠, 99.98% trace metals basis
Sigma-Aldrich
氟化钠, BioXtra, ≥99%
Sigma-Aldrich
2-(5-溴-2-吡啶偶氮)-5-二乙氨基酚, 97%