Solution Informationhelp
Enzyme: Dual specificity protein phosphatase 6
inhibitor: BDBM35435
substrate: n/a
 
Solution Type: Aqueous
pH at Preparation: n/a
Temp. Prep.: n/a
Comments: MKP-3 Assay Protocol Materials: 1. 10X Assay Buffer: 300 mM Tris, 750 mM NaCl, 10 mM EDTA, pH 7.0. Trizma 47.28 g, NaCl 43.83 g, EDTA 3.722 g, dH2O 800mL, pH to 7.0 with NaOH add dH2O to 1000mL 2. DTT: 200 mM Stock in -20 ◦C Freezer F17 3. BSA: 6.6% BSA Stock in -20 ◦C Freezer F17 4. OMFP Substrate: 5.0 uL/well ~ 100 uM final (2.5% DMSO). Add 1.05 mg OMFP from Freezer F17 per 0.5 mL of DMSO and sonicate in water bath ~ 5 min to make a 4.0 mM stock of OMFP (100% DMSO). Immediately prior to the assay dilute the 4.0 mM OMFP to 300 uM in dH2O and transfer 5 uL to each well of a 384-well of the assay plate ~ 100 uM OMFP final (2.5% DMSO). 5. MKP-3 Enzyme + Catalase 100 U/mL (Final): 5.0 uL/well ~ 125 or 250 ng/well MKP-3 + Catalase at 300 U/mL. Partially purified MKP-3 enzyme at 642, 1119 and 648 ug/mL for enzyme preparation I, II and III respectively was stored in 0.5 mL aliquots stored at -80 ◦C Dilute MKP-3 to the required protein concentration in 3X assay bu
 
 
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