Data Sheet EZH2 Homogeneous Assay Kit
Contents
1. 6044 Cornerstone Ct West Ste E San Diego CA 92121 Bioscience Tel 1 858 829 3082 Fax 1 858 481 8694 Email info bpsbioscience com Data Sheet EZH2 Homogeneous Assay Kit Catalog 52059 DESCRIPTION The EZH2 Homogeneous Assay Kit is designed to measure EZH2 activity for screening and profiling applications EZH2 is a histone methyltransferase that exhibits methylation activity toward H3 K27 The EZH2 Homogeneous Assay Kit comes in a convenient AlphaLISA format with histone substrate mixture his tagged primary antibody methylation assay buffer and purified EZH2 for 384 enzyme reactions The key to the EZH2 Homogeneous Assay Kit is a highly specific antibody that recognizes methylated substrate With this kit only three simple steps on a microtiter plate are required for methyltransferase detection First a sample containing EZH2 enzyme is incubated with the histone substrate for one hour Next acceptor beads and primary antibody are added then donor beads followed by reading the Alpha counts COMPONENTS Catalog Component Amount Storage Storage 51004 EZH2 100 ug 80 C Avoid freeze th aw cycles 52120 A_ 100 uM S adenosylmethionine 400 ul 80 C 52140H_ Primary antibody 8 5 pl 4 C Histone mixture his tagged 400 ul 80 C 52170 A_ 4x HMT assay buffer 2 3 ml 20 C 4x Detection buffer 2 2 ml 20 C MATERIALS REQUIRED BUT NOT SUPPLIED AlphaLISA anti rlgG accept2. R DIAGNOSTIC OR THERAPEUTIC USE To place your order please contact us by Phone 1 858 829 3082 Fax 1 858 481 8694 Or you can Email us at info bpsbioscience com Please visit our website at www bpsbioscience com 121008Final 6044 Cornerstone Ct West Ste E San Diego CA 92121 amp Bioscience Tel 1 858 829 3082 Fax 1 858 481 8694 Email info bpsbioscience com Example of Assay Results EZH2 activity ALPHA Screen 40000 E 30000 gt e 20000 5 lt m lt i 40000 0 0 50 100 150 200 250 EZH2 ng EZH2 enzyme activity measured using the EZH2 Homogeneous Assay Kit BPS Bioscience Cat 52059 Data shown is lot specific For lot specific information please contact BPS Bioscience Inc at info bpsbioscience com REFERENCES Dillon SC et al 2005 Genome Biology 6 227 RELATED PRODUCTS EZH1 EED SUZ12 Recombinant Protein 51006 50 ug EZH1 EED SUZ12 RbAp48 AEBP2 Complex Recomb 51007 50 ug EZH2 EED inactive Recombinant Protein 51002 20 ug EZH2 EED SUZ12 Recombinant Protein 51003 50 ug EZH2 EED SUZ12 RbAp48 AEBP2 Complex Recomb 51004 50 ug EZH2 Chemiluminescent Assay Kit 52009L 96 reactions AlphaScreen and AlphaLISA are registered trademarks of PerkinElmer Inc OUR PRODUCTS ARE FOR RESEARCH USE ONLY NOT FOR DIAGNOSTIC OR THERAPEUTIC USE To place your order please contact us by Phone 1 858 829 3082 Fax 1 858 481 8694 Or you can Email us at info bpsbio
3. g undiluted enzyme in aliquots at 80 C Note EZH2 is very sensitive to freeze thaw cycles Do not re use thawed aliquots or diluted enzyme 2 Dilute EZH2 in 1X HMT assay buffer at 100 ng ul Keep diluted enzyme on ice until use Discard any unused diluted enzyme after use 3 Using master mixes as much as possible add the following reagents to the microwells in duplicate Blank Substrate Positive Test Sample Control Control EZH2 100 ng ul S 2 5 pl 2 5 ul 2 5 pl 4x HMT assay buffer 2 2 5 ul 2 5 ul 2 5 pl 2 5 pl 100 uM S adenosylmethionine 7 pl 1 pl 1 ul Histone substrate 1 ul 1 ul 1 ul 1 ul Test Inhibitor Activator X pl HO 5 5 ul 4 ul 3 ul 3 X pl Total 10 wl 10 ul 10 wl 10 ul 4 Add the entire reaction mixture 10 pl to the well of a 384 well white plate Incubate at room temperature for 1 hour Step 2 Note Protect your samples from direct exposure to light 1 Dilute anti Rabbit Acceptor beads PerkinElmer 6760607C or AL104C 150 fold with 1x Detection buffer Add 5 ul per well Shake plate briefly 2 Dilute Primary antibody 8 400 fold with 1x Detection buffer Add 5 ul per well Shake plate Incubate 30 min at room temperature Step 3 1 Dilute AlphaScreen Nickel donor beads PE AS101D 100 fold with 1x Detection buffer Add 10 ul per well Incubate for 30 60 min at room temperature 2 Read Alpha counts OUR PRODUCTS ARE FOR RESEARCH USE ONLY NOT FO
4. or beads 5 mg ml PerkinElmer AL104C AlphaScreen Nickel donor beads 5 mg ml PerkinElmer AS101D Optiplate 384 PerkinElmer 6007290 AlphaScreen microplate reader APPLICATIONS Great for studying enzyme kinetics and HTS applications CONTRAINDICATIONS Green and blue dyes that absorb light in the AlphaScreen signal emission range 520 620 nm such as Trypan Blue Avoid the use of the potent singlet oxygen quenchers such as sodium azide NaN3 or metal ions Fe2 Fe3 Cu2 Zn2 and Ni2 The presence of gt 1 RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium MEM which lacks these components does not affect AlphaScreen assays STABILITY At least one year from date of receipt when stored as directed OUR PRODUCTS ARE FOR RESEARCH USE ONLY NOT FOR DIAGNOSTIC OR THERAPEUTIC USE To place your order please contact us by Phone 1 858 829 3082 Fax 1 858 481 8694 Or you can Email us at info bpsbioscience com Please visit our website at www bpsbioscience com 121008Final 6044 Cornerstone Ct West Ste E San Diego CA 92121 Tel 1 858 829 3082 Fax 1 858 481 8694 Email info bpsbioscience com Bioscience ASSAY PROTOCOL All samples and controls should be tested in duplicate Step 1 1 Thaw EZH2 on ice Upon first thaw briefly spin tube containing enzyme to recover full content of the tube Aliquot EZH2 enzyme into single use aliquots Store remainin
5. science com Please visit our website at www bpsbioscience com 121008Final
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