Data Sheet
Contents
1. Assay reagents Test sample Solvent Inhibitor control control Kinase Reaction Buffer 80 uL 80 uL 80 uL 10X Inhibitor or equivalent 10 uL Solvent forInhibitor 10 uL 10X Staurosporine 200 uM 10 pL Weel positive control 4 m units uL 10 uL 10 uL 10 pL or your enzyme fraction Kinase Reaction Buffer See Page 6 section Preparation of Working Solution Cat S 4400 See Page 4 section Materials Required but not Provided Even though final concentration of Staurosporine is 20 uM kinase activity of Weel isn t completely inhibited See Fig 2 Weel positive control Cat CY E1172 See Page 4 section Materials Required but not Provided Cat CY 1172 8 Version 120420 4 yy YCLEX User s Manual Weel Kinase Assay Inhibitor Screening Kit For Research Use Only Not for use in diagnostic procedures 1 Following the above table add the Reagents to each well of the microplate Finally initiate reaction by adding 10 uL of Weel positive control to each well and mixing thoroughly at room temperature Cover with plate sealer or lid and incubate at 30 C for 60 minutes 2 Follow the Standard Assay steps 5 12 page 6 7 Special considerations when measuring precise Weel activity In order to measure the activity of Weel correctly it is necessary to conduct the control experiment of Inhibitor control at least once for every experiment and ATP mi2. 0 5 10 15 20 25 30 35 40 Weel m units 100 ul reaction Fig 2 Effect of broad spectrum kinase inhibitor staurosporine on Weel activity 120 gt 100 80 60 40 Intensity of Conrol 20 0 0 001 0 010 0 100 1 000 10 000 100 000 Staurosporine Conc uM Cat CyY 1172 11 Version 120420 f cA p ay ycLex User s Manual U P Nn an N oO O Weel Kinase Assay Inhibitor Screening Kit For Research Use Only Not for use in diagnostic procedures References Nurse P Checkpoint pathways come of age Ce 91 865 867 1997 Watanabe N M Broome and T Hunter Regulation of the human WEE1Hu CDK tyrosine 15 kinase during the cell cycle EMBO J 14 1878 1891 1995 McGowan C H and P Russell Cell cycle regulation of human Weel EMBO J 14 2166 2175 1995 Mueller PR Coleman TR Kumagai A Dunphy WG Mytl a membrane associated inhibitory kinase that phosphorylates Cdc2 on both threonine 14 and tyrosine 15 Science 270 86 90 1995 Michael W M and J Newport Coupling of mitosis to the completion of S phase through Cdc34 mediated degradation of Weel Science 282 1886 1889 1998 Shi L W K Nishioka J Th ng E M Bradbury D W Litchfield and A H Greenberg Premature p34 activation required for apoptosis Science 263 1143 1145 1994 Leach S D C D Scatena C J Keefer H A Go
3. 5 12 page 6 7 Cat CyY 1172 Version 120420 pry Weel Kinase Assay Inhibitor Screening Kit ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Evaluation of Results Average the absorbance values for the Weel sample duplicates positive control and all experimental sample duplicate values when applicable When Weel positive control 40 m units assay is included as an internal control for the phosphorylation reaction the absorbance value should be greater than 1 5 with a background less than 0 3 2 For screening of purification chromatography fractions on graph paper plot the mean absorbance values for each of the samples on the Y axis versus the fraction number on the X axis to determine the location of the eluted purified Weel WwW For kinetic analysis on graph paper plot the mean absorbance values for eachjof the time points on the Y axis versus the time of each reaction minutes on the X axis Troubleshooting The CycLex Weel positive control Cat CY E1172 should b run in duplicate when a standard assay is being performed using the protocol described in the Detailed Protocol Incubation times or temperatures significantly different from those specified may give erroneous results 2 The reaction curve is nearly a straight line if the kinetics ofthe assay is of the first order Variations in the protocol can lead to non linearity of the curv
4. Substrate Reagent v Add 100 uL of Stop Solution v Measure absorbance at 450 nm Cat CyY 1172 3 Version 120420 pry Weel Kinase Assay Inhibitor Screening Kit ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Materials Provided All samples and standards should be assayed in duplicate The following components are supplied and are sufficient for the one 96 well microtiter plate kit Microplate One microplate supplied ready to use with 96 wells 12 strips of 8 wells in a foil zip lock bag with a desiccant pack Wells are coated with recombinant Cdc2 as a Weel substrate 10X Wash Buffer One bottle containing 100 mL of 10X buffer containing 2 Tween 20 Kinase Buffer One bottle containing 20 mL of 1X buffer used for Kinase Reaction Buffer and sample dilution 20X ATP Lyophilized ATP Nay salt Anti Phospho Tyrosine Monoclonal Antibody PY 39 One vial containing 12 mL of anti phospho tyrosine Ready to use HRP conjugated Anti mouse IgG One vial containing 12 mb of HRP horseradish peroxidase conjugated anti mouse IgG Ready to use Substrate Reagent One bottle containing 20 mL ofthe chromogenic substrate tetra methylbenzidine TMB Ready to use Stop Solution One bottle containing 20 mL of 1 N H2SOx Ready to use Materials Required but not Provided e Weel positive control Available from CycLex Weel positive control Cat CY E1172 One vial contains 8 uni
5. CAUTION sSulfuric Acid is a strong acid Wear disposable gloves and eye protection when handling Stop Solution Cat CyY 1172 5 Version 120420 yy os yclex Detailed Protocol User s Manual Weel Kinase Assay Inhibitor Screening Kit For Research Use Only Not for use in diagnostic procedures The CycLex Weel kinase Assay Inhibitor Screening Kit is provided with removable strips of wells so the assay can be carried out on separate occasions using only the number of strips required for the particular determination Since conditions may vary running an aliquot of the appropriate Weel positive control Cat CY E1172 separately available from CycLex should be included in each assay Disposable pipette tips and reagent troughs should be used for all transfers to avoid cross contamination of reagents or samples Preparation of Working Solution 1 Prepare a working solution of Wash Buffer by adding 100 mL of the 10X Wash Buffer provided to 900 mL of ddH20 Mix well Store at 4 C for two weeks or 20 C for long term storage 2 Prepare 20X ATP Solution by adding 0 8 mL of ddH O to the vial of 20X ATP provided lyophilized Mix gently until dissolved The final concentration of the 20X ATP Solution should be 2 5 mM Store the solution in small aliquots e g 100 uL at 20 G 3 Prepare Kinase Reaction Buffer by mixing following reagents 96 assays 10 assays 1 assay Kinase Buffer provide
6. Weel kinase 2 Detecting the effects of pharmacological agents on Weel kinase activity in vitro This assay kit is for research use only and not for use in diagnostic or therapeutic procedures Storage e Upon receipt Store all components at 4 C Don t expose reagents to excessive light Cat CyY 1172 1 Version 120420 Weel Kinase Assay Inhibitor Screening Kit wy cA ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Introduction Weel kinase negatively regulates entry into mitosis by catalyzing the inhibitory tyrosine phosphorylation of Cdc2 cyclin B kinase 1 Antibody depletion experiments demonstrate that Weel accounts for most of the activity that phosphorylates Cdc2 on Tyr15 within the ATP binding pocket of the Cdc2 catalytic subunit in an in vitro assay of HeLa cell lysates 2 3 hence it is likely to have an important role in the mitotic control of human cells While Myt1 is responsible for phosphorylation of another inhibitory threonine Thr14 of Cdc2 cyclin B kinase 4 Weel kinase activity is strongly suppressed during M phase suggesting that negative regulation of Weel could be part of the mechanism by which activation of Cdc2 cyclin B kinases promoted during the G2 M transition Weel activity increased during S and G2 phases in parallel with the level of protein its activity decreased at M phase when Weel became transiently hyperphosphorylated 2 3 In addition a decre
7. ase in Weel protein level was observed at M G1 phase 2 3 Apparently the hyperphosphorylation and degradation in combination caused inactivation of Weel at M phase and the following G1 phase 4 These results suggest that the activity of Weel is regulated by phosphorylation and proteolytic degradation and that Weel plays a role in inhibiting mitosis before M phase by phosphorylating Cdc2 cyclin B kinase 1 3 5 Mammalian cells undergo cell cycle arrest in response to DNA damage due to the existence of multiple checkpoint response mechanisms One such checkpoint pathway operating at the G1 phase is frequently lost in cancer cells due to mutation of the p53 tumofsuppressor gene However cancer cells often arrest at the G2 phase upon DNA damage due to activation of another checkpoint pathway that prevents the activation Cdc2 kinase The kinases Weel and Chkl are key regulators of this G2 checkpoint 6 10 which act directly or indirectly to inhibit Cdc2 activity Inhibition of Weel and Chk1 sensitized only cancer cells which lost G1 phase checkpointto DNA damage agents induced apoptosis These data support the attractiveness of Weel as well as Chk1 is as molecular targets for abrogating the G2 DNA damage checkpoint arrest a situation that may selectively sensitize p53 deficient tumor cells to radiation or chemotherapy treatment Measurement of Weel activity The protocol generally regarded as most sensitive for the quantitative measurement of W
8. c Assays 1 Remove the appropriate number of microtiter wells from the foil pouch and place them into the well holder Return any unused wells to the foil pouch refold seal with tape and store at 4 C 2 Prepare all samples diluted with Kinase Buffer as needed All samples should be assayed in duplicate 3 Duplicate wells containing 10 uL of Weel positive control 40 m units should be included in each assay as a positive control for phosphorylation 4 lt Begin the kinase reaction by addition of 90 uL of Kinase Reaction Buffer in duplicate per well in timed intervals suggested interval is 5 minutes but should be individually determined for each system After the final addition cover with plate sealer or lid and incubate at 30 C for 40 minutes 5 Stop the reaction by flicking out the contents Alternatively the reaction may be terminated by the addition of 150 uL 0 1 M Na EDTA pH 8 0 to each well Cat CY 1172 7 Version 120420 Weel Kinase Assay Inhibitor Screening Kit 4 gt ycLex User s Manual For Research Use Only Not for use in diagnostic procedures 6 Wash wells four times with Wash Buffer making sure each well is filled completely Remove residual Wash Buffer by gentle tapping or aspiration 7 Pipette 100 uL of Anti Phospho Tyrosine Monoclonal Antibody PY 39 into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 60 minutes Discard any unused antibody after us
9. d 9 5 mL 950 pL 95 uL 20X ATP provided 0 5 mL 50 uL 5 pL Total 10 mL 1000 uL 100 pL You will need 80 90 uL of Kinase Reaction Buffer per assay well Mix well Discard any unused Kinase Reaction Buffer after use Standard Assay 1 Remove the appropriate number of microtiter wells from the foil pouch and place them into the well holder Return any unusedwells to the foil pouch refold seal with tape and store at 4 C 2 Prepare all samples diluted with Kinase Buffer as needed All samples should be assayed in duplicate 3 Duplicate wells containing 10 uL of Weel positive control 40 m units should be included in each assay asia positive control for phosphorylation 4 Begin the kinase reaction by addition of 90 uL Kinase Reaction Buffer per well cover with plate sealer or lid and incubate at 30 C for 60 minutes 5 Wash wells four times with Wash Buffer making sure each well is filled completely Remove residual Wash Buffer by gentle tapping or aspiration 6 Pipette 100 uL of Anti Phospho Tyrosine Monoclonal Antibody PY 39 into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 60 minutes Discard any unused antibody after use Cat CyY 1172 Version 120420 Weel Kinase Assay Inhibitor Screening Kit wy cA ycLex User s Manual For Research Use Only Not for use in diagnostic procedures 7 Wash wells four times as same as in step 5 8 Pipette 100 uL
10. e 8 Wash wells four times as same as in step 5 9 Pipette 100 uL of HRP conjugated Anti mouse IgG into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 60 minutes Discard anyetinused conjugate after use 10 Wash wells five times as same as in step 5 11 Add 100 pL of Substrate Reagent to each well and incubate at room temperature for 5 15 minutes 12 Add 100 uL of Stop Solution to each well in the same order as the previously added Substrate Reagent 13 Measure absorbance in each well using a spectorphotometri plate reader at dual wavelengths of 450 540 nm Dual wavelengths of 450 550 or 450 595 nm can also be used Read the plate at 450 nm if only a single wavelength can be used Wells must be read within 30 minutes of adding the Stop Solution Recommendations Special considerations when screening activators or inhibitors In order to estimate the inhibitory effect on individual Weel activity in the test chemicals correctly it is necessary to conduct the control experiment of Solvent control at least once for every experiment and Inhibitor control at least once for the first experiment in addition to Test sample as indicated in the following table When test chemicals cause an inhibitory effect on Weel activity the level of A450 is weakened as compared with Solvent control The high level of A450 is not observed in Inhibitor control usually A450 lt 0 4
11. e as can assay kinetics of other than first order For a non linear curve point to point or quadratic curve fit methods should be used U Poor duplicates accompanied by elevated values for wells containing no sample indicate insufficient washing If all instructions in the Detailed Protocol were followed accurately such results indicate a need for washer maintenance 4 Overall low signal may indicate that desiccation of the plate has occurred between the final wash and addition of Substrate Reagent Do not allowsthe plate to dry out Add Substrate Reagent immediately after wash Reagent Stability All of the reagents included in the CycLex Research Product CycLex Weel kinase Assay Inhibitor Screening Kit have been tested for stability Reagents should not be used beyond the stated expiration date Upon receipt kit reagents should be stored at 4 C except the ATP must be stored at 20 C Coated assay plates should be stored in the original foil bag sealed by the zip lock and containing a desiccant pack For research use only not for use in diagnostic or therapeutic procedures Cat CyY 1172 10 Version 120420 P Weel Kinase Assay Inhibitor Screening Kit PA c ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Example of Test Results Fig 1 Dose dependency of recombinant Weel enzyme reaction 3 0 A450 0 0 tt ht tt BED CTO CET EE EEN GEN EE ED AE Tt
12. eel activity involves incubation of the Weel sample with substrate either a natural or synthetic polypeptide such as weel tide Cdc2 peptide in the_presence of Mg and P labeled ATP The reaction is terminated by spotting a sample onto a phospho cellulose P81 filter paper disc followed by washing extensively to remove unincorporated radiolabel and the radioactivity counted While sensitive this method is labor intensive generates hazardous radioactive waste and depends on a radioisotope of short half life It is particularly unsuitable when kinase assays are only performed on an infrequent basis The CycLex Weel kinase Assay Inhibitor Screening Kit uses peroxidase coupled anti Phospho Tyrosine monoclonal antibody as a reporter molecule in a 96 well ELISA format This assay provides a non isotopic sensitive and specific method to measure the activities of Weel kinase Cat CyY 1172 2 Version 120420 f c p Weel Kinase Assay Inhibitor Screening Kit ay ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Principle of the Assay The CycLex Research Product CycLex Wee1 kinase Assay Inhibitor Screening Kit is a single site semi quantitative immunoassay for Weel activity Plates are pre coated with a substrate corresponding to recombinant Cdc2 which contains tyrosine15 residues that can be phosphorylated by Weel The detector antibody specifically detects only the phosphorylated form of tyrosi
13. ne15 residue on Cdc2 The CycLex Weel kinase Assay Inhibitor Screening Kit can be used to study the kinetics fia purified or partially purified Weel as well as to screening Weel inhibitor To perform the test the sample is diluted in Kinase Buffer pipetted into the wells and allowed to phosphorylate the bound substrate following the addition of Mg and ATP The amount of phosphorylated substrate is measured by binding it with a PY 39 an anti Phospho Tyrosine monoclonal antibody followed by binding with horseradish peroxidase conjugated anti mouse IgG which then catalyzes the conversion of the chromogenic substrate tetra methylbenzidine TMB from a colorless solution to a blue solution or yellow after the addition of stopping reagent The color is quantitated by spectrophotometry and reflects the relative amount of Weel activity in the sample For kinetic analysis the Weel containing sample is added to the wells in a similar fashion and at varying times the reaction is stopped by the addition of the chelator sodium ethylenediaminetetraacetate EDTA and the amount of phosphorylated substrate determined as before Summary of Procedure Add 100 uL of sample to the wells y Incubate for 60 min at 30 C Wash the wells Add 100 uL of Anti Phospho Tyrosine antibody PY 39 Yy Incubate for 60 min at room temp Wash the wells Add 100uL of HRP conjugated anti mouse IgG y Incubate for 60 min at room temp Wash the wells v Add 100 uL of
14. nus control at least once for the first experiment in addition to No enzyme control as indicated in the following table Although the level of A450 increases in Test sample when Weel enzyme activity is in the sample the high level of A450 is not observed in Inhibitor control ATP minus control and No enzyme control Assay reagents Test Inhibitor ATP minus Positive No enzyme Sample control control control control Kinase Reaction buffer 90 pL 80 uL 90 pL 90 pL Kinase Buffer ATP minus 90 uL 10X Staurosporine 200 uM 10 uL Your enzyme fraction 10 uL 10 uL 10 pL Weel positive control 4 m units uL 10 uL Buffer 10 uL Kinase Reaction Buffer See Page 6 section Preparation of Working Solution Cat S 4400 See Page 4 section Materials Required but not Provided Even though final concentration of Staurosporine is 20 uM kinase activity of Weel isn t completely inhibited See Fig 2 Weel positive control Cat CY E1172 See Page 4 sectiom Materials Required but not Provided 1 Following the above table add the Reagents toveach well of the microplate Finally initiate the reaction by adding 10 uL of Your enzyme fraction or Buffer to each well and mixing thoroughly at room temperature Cover with plate sealer or lid and incubate at 30 C for 60 minutes 2 Follow the Standard Assay steps
15. odman S Y Song L Yang and J A Pietenpol Negative regulation of Weel expression and Cdc2 phosphorylation during p53 mediated growth arrest and apoptosis Cancer Res 58 3231 3236 1998 Yuan H Xie Y M and Chen I S Depletion of Wee I kinase is necessary for both human immunodeficiency virus type 1 Vpr and gamma irradiation inducedapoptosis J Virol 77 3 2063 2070 2003 Heald R M McLoughlin and F McKeon Human weel maintains mitotic timing by protecting the nucleus from cytoplasmically activated Cde2 kinase Cell 74 463 474 1993 10 Jackson JR Gilmartin A Imburgia C Winkler JD Marshall LA Roshak A An indolocarbazole inhibitor of human checkpoint kinase Chk1 abrogates cell cycle arrest caused by DNA damage Cancer Res 60 3 566 72 2000 Cat CyY 1172 12 Version 120420 ry Weel Kinase Assay Inhibitor Screening Kit rb y 3X User s Manual For Research Use Only Not for use in diagnostic procedures Related Products Weel Positive control Cat CY E1172 CycLex Protein Phosphatase Cdc25A Fluorometric Assay Kit Cat CY 1352 CycLex Protein Phosphatase Cdc25B Fluorometric Assay Kit Cat CY 1353 CycLex Protein Phosphatase Cdc25C Fluorometric Assay Kit Cat CY 1354 CycLex Protein Phosphatase Cdc25 Combo Fluorometric Assay Kit Cat CY 1355 PRODUCED BY CycLex Co Ltd 1063 103 Terasawaoka Ina Nagano 396 0002 Japan Fax 81 265 76 7618 e mail info cyclex co jp URL ht
16. of HRP conjugated Anti mouse IgG into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 60 minutes Discard any unused conjugate after use 9 Wash wells five times as same as in step 5 10 Add 100 uL of Substrate Reagent to each well and incubate at room temperature ca 25 C for 5 15 minutes 11 Add 100 uL of Stop Solution to each well in the same order as the previously added Substrate Reagent 12 Measure absorbance in each well using a spectrophotometric plate reader at dual wavelengths of 450 540 nm Dual wavelengths of 450 550 or 450 595 nm can also bemused Read the plate at 450 nm if only a single wavelength can be used Wells must be read within 30 minutes of adding the Stop Solution Note 1 Complete removal of liquid at each step is essential to good performance After the last wash remove any remaining Wash Buffer by aspifating or decanting Invert the plate and blot it against clean paper towels Note 2 Reliable signals are obtained when either O D values do not exceed 0 3 for the blank no enzyme control or 1 5 for the Weel positive control Note 3 If the microplate reader is not capable of reading absorbance greater than the absorbance of the Weel positive control perform a Second reading at 405 nm A new O D values measured at 405 nm is used to determine Weel activity of off scale samples The readings at 405 nm should not replace the on scale readings at 450 nm Kineti
17. ot expose reagents to excessive light Avoid freeze thaw cycles Allow all the components to come to room temperature before use e All microplate strips that are not immediately required should be returned to the zip lock pouch which must be carefully resealed to avoid moisture absorption e Do not use kit components beyond the indicated kit expiration date e Use only the microtiter wells provided with the kit e Rinse all detergent residue from glassware e Use deionized water of the highest quality e Do not mix reagents from different kits e The buffers and reagents used in this kit contain either sodium Kathon CG as preservatives Care should be taken to avoid direct contact with these reagents e Do not mouth pipette or ingest any of the reagents e Do not smoke eat or drink when performing the assay or in areas where samples or reagents are handled e Dispose of tetra methylbenzidine TMB containing solutions in compliance with local regulations e Avoid contact with Substrate Solution which contains hydrogen peroxide e Avoid contact with Stop Solution which contains Sulfuric Acid e In case of contact with the Stop Solution and the Substrate Solution wash skin thoroughly with water and seek medical attention when necessary e Biological samples may be contaminated with infectious agents Do not ingest expose to open wounds or breathe aerosols Wear protective gloves and dispose of biological samples properly e
18. pry Weel Kinase Assay Inhibitor Screening Kit cA ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Non Radioisotopic Kit for Measuring Weel Kinase Activity CycLex Weel Kinase Assay Inhibitor Screening Kit Cat CY 1172 Tntend d US isscscacsdicdavasenadanslenciasieateexaeietiann 1 PMI AR SE T 1 MAIO saniser iea 2 Principle of the ASSAY ssccsssisdeeaseescanesstvencesed 3 Materials Provided cccccccceesseeeeeeeseeees 4 Materials Required but not Provided 4 Precautions and Recommendation 5 Detailed Protocol cccccsccccessseceeeesseeeees 6 9 Evaluation of Results cccccceccceeeseeees 10 Assay Characteristics sas iictesveccssdusansineete Sines 10 Troubleshooting sxxscsnecescha Hsscasadoinesourecnehasivs 10 Reagent Stability i xsiccccsstecessdesdehesntdeccevinndateas 10 Example of Test Results ceceeeeseeeees 11 Referentes ennienni i 12 Related Products wiscssssisscstasrnecsctecngeme 13 Intended Use The CycLex Research Product CyeLex Weel Kinase Assay Inhibitor Screening Kit is designed to measure the activities of purified Weel for the rapid and sensitive evaluation of inhibitors or activators The phospho tyrosine specific monoclonal antibody used in this assay kit has been demonstrated to recognize the phospho tyrosing 15 residue in Cdc2 which is phosphorylated by Wee1 Applications of this kit include 1 Screening inhibitors or activators of
19. tp www cyclex co jp CycLex CircuLex products are supplied for research use only CycLex CircuLex products and components thereof may not be resold modified for resale or used to manufacture commercial products without prior written approval from CycLex Co Ltd To inquire about licensing for such commercial use please contact us via email Cat CyY 1172 13 Version 120420
20. ts 200 uL of Weel enzyme The Positive control should be added to the first well at 40 m units well For instance diluted positive control 1 10 use 10 uL for 1 assay Unused Weel enzyme should be stored in aliquots at below 70 C e Staurosporine 200 uM Staurosporine is available from Sigma Cat S 4400 1 mM stock solution DMSO diluted 1 5 with Kinase Buffer Even though final concentration of Staurosporine is 20 uM kinase activity of Weel isit completely inhibited See Fig 2 Pipettors 2 20 uL 20 200 uL and 200 1000 uL precision pipettors with disposable tips Precision repeating pipettor e Wash bottle or multichannel dispenser for plate washing e Microcentrifuge andtubes for sample preparation e Vortex mixer e Plate reader capable of measuring absorbance in 96 well plates at dual wavelengths of 450 nm 540 nm Dual wavelengths of 450 550 or 450 595 nm can also be used The plate can also be read at a single wavelength of 450 nm which will give a somewhat higher reading 500 or 1000 mL graduated cylinder Reagent reservoirs Deionized water of the highest quality Disposable paper towels Cat CyY 1172 4 Version 120420 Weel Kinase Assay Inhibitor Screening Kit 4 gt ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Precautions and Recommendations Store the Weel enzyme at below 70 C and the ATP at 20 C when not in use Store all other components at 4 C Do n
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