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Data Sheet LSD1 Chemiluminescent Assay Kit

1. C Dall Aglio A Fraaije MW Battaglioli E and Mattevi A J Biol Chem 2006 281 46 35289 95 2 Zhou M Diwu Z Panchuk Voloshina N and Haugland RP Anal Biochem 1997 253 2 162 8 ASSAY PROTOCOL All samples and controls should be tested in duplicate Step 1 1 Rehydrate the microwells by adding 200 ul of TBST buffer 1 x TBS pH 8 0 containing 0 05 Tween 20 to every well Incubate 15 minutes at room temperature Tap the plate onto clean paper towels to remove liquid Thaw LSD1 on ice Upon first thaw briefly spin tube containing enzyme to recover full content of the tube Aliquot LSD1 enzyme into single use aliquots Store remaining undiluted enzyme in aliquots at 80 C Note LSD1 is sensitive to freeze thaw cycles Do not re use thawed aliquots or diluted enzyme Add 10 ul 3x LSD1 assay buffer 2 15 ul distilled H20 to each well Dilute LSD1 in 1X LSD1 assay buffer 2 at 5 ng ul Keep diluted enzyme on ice until use Discard any unused diluted enzyme after use Add 5 ul of Inhibitor solution of each well designated Test Inhibitor For the Positive Control and Blank add 5 ul of the same solution without inhibitor Inhibitor buffer Add 20 ul of 1X LSD1 assay buffer 2 to the well designated Blank OUR PRODUCTS ARE FOR RESEARCH USE ONLY NOT FOR DIAGNOSTIC OR THERAPEUTIC USE 130227 To place your order please contact us by Phone 1 858 829 3082 Fax 1 858 481 8694 Or you can Email us
2. chemiluminescence reader COMPONENTS Catalog Component Amount Storage 50100 LSD1 10 ug 80 C 52140J Primary antibody 10 25 ul 80 C 52130H Secondary HRP labeled antibody 1 10 ul 80 C 3x LSD1 assay buffer 2 3 ml 20 C Avoid 52100 Blocking buffer 50 ml 4 C freeze HRP chemiluminescent substrate A 6ml 4 C thaw transparent bottle cycles HRP chemiluminescent substrate B 6 ml 4 C brown bottle Black microplate precoated with histone 1 4 C substrate MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED TBST buffer 1 x TBS pH 8 0 containing 0 05 Tween20 Luminometer or fluorescent microplate reader capable of reading chemiluminescence Adjustable micropipettor and sterile tips Rotating or rocker platform APPLICATIONS Great for studying enzyme kinetics and HTS applications 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 130227 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 CONTRAINDICATIONS DMSO gt 1 strong acids or bases ionic detergents high salt STABILITY One year from date of receipt when stored as directed REFERENCES 1 Forneris F Binda
3. 0103 20 ug JMJD2B recombinant protein 50104 20 ug JMJD2C recombinant protein 50105 20 ug JMJD2E recombinant protein 50118 20 ug 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 130227
4. 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 LSD1 Chemiluminescent Assay Kit Catalog 50109 DESCRIPTION The LSD1 Chemiluminescent Assay Kit is designed to directly measure activity of human lysine specific demethylase LSD1 enzymes containing LSD1 for screening and profiling applications LSD1 is a chromatin modifying enzyme that specifically removes methyl groups from mono and di methylated Lys of histone H3 LSD1 is a critical component of transcriptional regulation via epigenetic histone modifications and is therefore a potential target for drug development The LSD1 Chemiluminescent Assay Kit comes in a convenient format with a 96 well plate precoated with the methylated histone H3 peptide substrate primary antibody the secondary HRP labeled antibody demethylase assay buffer and purified LSD1 for 100 enzyme reactions The key to the LSD1 Chemiluminescent Assay Kit is a highly specific antibody that recognizes demethylated substrate With this kit only three simple steps on a microtiter plate are required for methyltransferase detection First a sample containing LSD1 enzyme is incubated with a sample containing assay buffer Next primary antibody is added Finally the plate is treated with an HRP labeled secondary antibody followed by addition of the HRP substrate to produce chemiluminescence that can then be measured using a
5. at info bpsbioscience com Please visit our website at www bpsbioscience com 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 Positive Test Blank Control Inhibitor 3x LSD1 assay buffer 2 10 ul 10 ul 10 ul H2O 15 ul 15 ul 15 ul Inhibitor 5 ul Inhibitor buffer no inhibitor 5 ul 5 ul 1x LSD1 assay buffer 2 20 ul LSD1 5 ng ul 20 ul 20 ul Total 50 ul 50 ul 50 ul 7 Initiate reaction by adding 20 ul of diluted LSD1 enzyme to the wells designated Positive Control and Test Inhibitor Incubate at room temperature for 30 45 min 8 Remove the supernatant from the wells and wash the plate three times with 200 ul TBST buffer Blot dry onto clean paper towels 9 Add 100 ul of Blocking buffer to every well Shake on a rotating platform for 10 min Remove supernatant as described above Step 2 1 Dilute Primary antibody 10 400 fold with Blocking buffer 2 Add 100 ul per well Incubate 1 hour at room temperature with slow shaking 3 Remove the supernatant from the wells and wash plate three times with 200 ul TBST buffer and incubate in Blocking buffer as described in steps 1 8 and 1 9 Step 3 1 Dilute Secondary HRP labeled antibody 1 1 000 fold with Blocking buffer 2 Add 100 ul per well Incubate for 30 minutes at room temperature with slow shaking 3 Remove the s
6. d type endpoint Sensitivity may be adjusted based on luminescence of a control without enzyme typically we set this value as 100 when using Synergy 2 plate reader EXAMPLE OF ASSAY RESULTS LSD1 activity 600 500 400 300 Luminescence 200 100 0 0 10 20 30 40 50 60 70 80 90 100 110 LSD1 ng LSD1 enzyme activity measured using the LSD1 Chemiluminescent Assay Kit BPS Bioscience 50109 Data shown is lot specific For lot specific information please contact BPS Bioscience Inc at info bpsbioscience com 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 130227 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 RELATED PRODUCTS LSD1 recombinant protein 50100 50 ug LSD1 Fluorescent Assay Kit 50106 96 reactions LSD1 Fluorescent Assay Kit 50107 384 reactions LSD1 Homogeneous Assay Kit 50108 384 reactions LSD1 substrate 50101 500 ul JMJD2A Homogeneous Assay Kit 50413 384 reactions JMJD2B Homogeneous Assay Kit 50414 384 reactions JMJD2C Homogeneous Assay Kit 50415 384 reactions JMJD2E Homogeneous Assay Kit 50417 384 reactions JMJD2C Assay Kit Chemiluminescent 50405 96 reactions JMJD2A recombinant protein 5
7. upernatant from the wells and wash plate three times with 200 ul TBST buffer and incubate in Blocking buffer as described in steps 1 8 and 1 9 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 130227 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 4 Just before use mix on ice 50 ul HRP chemiluminescent substrate A and 50 ul HRP chemiluminescent substrate B and add 100 ul per well Discard any unused chemiluminescent reagent after use 5 Immediately read sample in a luminometer or microtiter plate reader capable of reading chemiluminescence Reading Chemiluminescence Chemiluminescence is the emission of light luminescence which results from a chemical reaction The detection of chemiluminescence requires no wavenlength selection because the method used is emission photometry and is not emission spectrophotometry To properly read chemiluminescence make sure you are using your plate reader in a LUMINESCENCE mode Typical integration time is 1 second delay after plate movement is 100 msec Make sure you don t have filter when emit the light Synergy 2 BioTek use hole position on filter wheel Optics position Top Rea

Data Sheet LSD1 Chemiluminescent Assay Kit

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