di-methylated K4_H3 peptide substrate
Catalog #: 50101
 
SIZE: 500 μl
 
DESCRIPTION: Histone H3 peptide methylated at Lysine 4, substrate for LSD1
 
PURITY: >90% by HPLC

MW: 2283
 
APPLICATIONS: Study enzyme kinetics, and screen small molecular inhibitors of
LSD1 for drug discovery and HTS applications
 
SUPPLIED AS: water solution
 
QUANTITY: 500 μl @200 μM
 
STORAGE: -20°C or -70°C

Assay Protocol:
 
Materials: 2x assay buffer, 200 μM methylated peptide substrate (BPS catalog number
50101), LSD1 (BPS catalog number 50100), Amplex Red reagent, peroxidase
 
Step 1: 
 
1) Add the entire reaction mixture to a 96 well plate:
 
20 μl LSD1 (10-15 ng/μl)
5 μl of 200 μM methylated peptide substrate
25 μl of 2x LSD1 assay buffer 
 
2) Incubate for 15-30 min at room temperature. 
 
Step 2: 
 
1) Immediately before use: add 50 μl of 10 mM Amplex Red stock and 100 μl of 10 U/ml
peroxidase to 4.85 ml of 1x LSD1 buffer, mix. 
2) Add 50 μl of  mixture to each well. Mix well.  
3) fluorescence at λexc=530 nm and λem=590 nm.