Small G-Protein Assay Kit

종류 및 특징
• RhoGEF exchange assay Biochem Kit (Cat# BK100)
  • GDP에서 GTP상으로 변할때 생성되는 guanine nucleotide exchange factor (GEF)의 양을 측정할 수 있습니다.
  • human Cdc42, Rac1, RhoA GTPases, GEF positive control인 GEF domain of Dbs뿐만 아니라 최적화된 시약이 제공됩니다.
  • 제품의 구성:
    • Exchange Buffer
    • His-tagged Cdc42 protein (Cat. # CD01)
    • His-tagged Rac1 protein(Cat. # RC01)
    • His-tagged RhoA protein (Cat. # RH01)
    • His-tagged hDbs protein (Cat. # GE01)
    • 384-well black half area round bottom plate
    • 96-well black half area flat bottom plate
    • Detailed instructions and troubleshooting manual
  • 적용
    • Determination of the activity and GTPase specificity of uncharacterized GEFs.
    • Biochemical characterization of small GTPases and their associated GEFs.
    • Examination of the regulation of GEF activity by different cofactors or protein domains.
    • Screen the mutant protein of either GEFs or GTPases for activity and substrate specificity.
    • Identification of GEF inhibitors in HTS (high throughput screen) format. Please inquire for significant discounts on large quantities of any reagents in this kit
  • 사용예
    • The exchange activity of Dbs on Cdc42, RhoA and Rac1 was measured in 96-well and 384-well format (Fig 1 & 2)
    
    Figure 1. Dbs exchange activity for Cdc42, RhoA and Rac1 in 96-well half area plate format
    
    Figure 2. Dbs exchange activity for Cdc42, RhoA and Rac1 in 384-well plate format
  • Product Citations
    • John, J., Sohmen, R., Feuerstein, J., Linke, R., Wittinghofer, A., and Goody, R. S. (1990) Kinetics of interaction of nucleotides with nucleotide-free H-ras p21. Biochemistry 29, 6058-6065.
    • Leonard, D. A., Evans, T., Hart, M., Cerione, R. A., and Manor, D. (1994) Investigation of the GTP-binding/GTPase cycle of Cdc42Hs using fluorescence spectroscopy. Biochemistry 33, 12323-12328.
• RhoGAP assay Biochem Kit (Cat# BK105)
  • GAP assay kit는 GTP에서 GDP + Pi로의 가수분해에 의해 생겨나는 inorganic phosphate (Pi)의 양을 측정합니다.
  • kit에는 GTPases, RhoGAP positive control, 최적화된 버퍼와 phosphate detection 시약이 포함되어 있습니다.
  • 제품의 구성:
    • Reaction Buffer
    • His-tagged Cdc42 protein (Cat. # CD01)
    • His-tagged Rac1 protein(Cat. # RC01)
    • His-tagged RhoA protein (Cat. # RH01)
    • His-tagged Ras protein (Cat. # RS01)
    • GST-tagged p50RhoGAP, catalytic domain (Cat. # GAS01)
    • GTP stock (Cat. # BST06)
    • CytoPhos™ reagent
    • 96-well clear half area flat bottom plate
    • Detailed instructions and troubleshooting manual
  • 적용
    • Determination of the activity and GTPase specificity of uncharacterized GAPs.
    • Biochemical characterization of small GTPases and their associated GAPs.
    • Examination of the regulation of GAP activity by different cofactors or protein domains.
    • Screen the mutant protein of either GAPs or GTPases for activity and substrate specificity.
    • Identification of GAP inhibitors in HTS (high throughput screen) format. Please inquire for significant discounts on large quantities of any reagents in this kit.
  • 사용예
    • The GTPase activity of RhoA with and without the addition of the catalytic domain of p50RhoGAP was tested (Fig 1).
    
    Figure 1. RhoGAP activity measured as GTP hydrolysis by RhoA protein. Each reaction contained reaction buffer + GTP with the addition of RhoA alone (RhoA), RhoGAP alone (GAP), or RhoA + RhoGAP (RhoA + GAP). Reactions were incubated at 37°C for 20 min. Phosphate generated by hydrolysis of GTP was measured by the addition of CytoPhos™ reagent and reading of absorbance at 650 nm.