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CY-1170

1. A 0 50 100 150 200 Heparin ng ml C CY 1170 12 Version 141117 CK2 KinaseAssay Inhibitor Screening Kit jde Urs i GA y gt X ser s Manu For Research Use Only Not for use in diagnostic procedures References j Lozeman F J Litchfield D W Piening C Takio K Walsh K A and Krebs E G 1990 Isolation and characterization of human cDNA clones encoding the a and a subunits of CK2 Biochemistry 29 8436 8447 2 Litchfield D W Lozeman FEJ Piening C Sommercorn J Takio K Walsh K A and Krebs E G 1990 Subunit structure of CK2 from bovine testis demonstration that the a and a subunits are distinct polypeptides J Biol Chem 265 7638 7644 3 Maridor G Park W Krek W and Nigg E A 1991 CK2 cDNA sequen es developmental expression and tissue distribution of mRNAs for a a and b subunits of the chicken enzyme J Biol Chem 266 2362 2368 4 Munstermann U Fritz G Seitz G Lu Y P Schneider H R and Issinger O G 1990 CK2 is elevated in solid human tumours and rapidly proliferating non neoplasti tissue Eur J Biochem 189 251 257 Nn Pepperkok R Lorenz P Ansorge W and Pyerin W 1994 CK2 1s required for transition of G0 G1 early G1 and GI S phases of the cell cycle J Biol Chem 269 6986 699 6 Landesman Bollag E Romieu Mourez R Song D H Sonenshein GE Cardiff R D and Seldin D C 2001 Protein kinas
2. Screening inhibitors or activators ofCK2 2 Detecting the effects of pharma ological agents on CK2 activity This assay kit is for research use only and not for use in diagnostic or therapeutic procedures Storage e Upon receipt store allseomponents at 4 C e Don t expose reagents to excessive light Cat CY 1170 1 Version 141117 CK2 KinaseAssay Inhibitor Screening Kit jde Urs i GA y gt X ser s Manu For Research Use Only Not for use in diagnostic procedures Introduction Protein kinase Casein kinase II CK2 is a ubiquitous and pleiotropic seryl threonyl protein kinase which appears to interact with different signaling pathways and therefore represents the prototype ofa multifunctional protein kinase The holoenzyme is generally composed of two catalytic alpha and or alpha and two regulatory beta subunits but the free alpha alpha subunits are catalyfieallyhactive by themselves Although the beta subunits deeply affect many properties of CK2 both the isolated catalytic subunits and the holoenzyme are constitutively active The enzyme is highly expressed in most cancers and this higher expression has been tentatively correlated with the involvement of CK in the promotion of specific phases of the cell cycle Unlike the majority of protein kinases which are tightly regulated enzymes CK2 is endowed with high constitutive activity a feature thatiis suspected to underlie its oncogenic potentia
3. 7 Cat CY 1170 Version 141117 y AA CK2 KinaseAssay Inhibitor Screening Kit User s Manual For Research Use Only Not for use in diagnostic procedures Special considerations when measuring precise CK2 activity In order to measure the activity of CK2 correctly it is necessary to conduct the control experiment of Inhibitor control at least once for every experiment and ATP minus control at least once for the first experiment in addition to No enzyme control as indicated in the following table Although the level ofi A450 increases in Test sample when CK2 enzyme activity is in the sample the high level of A450 is not observed in Inhibitor control ATP minus control and No enzyme control Inhibitor ATP minus Positive No enzyme Assay reagents Test Sample control control control control Kinase Reaction Buffer 90 pL 80 uL 90 pL 90 uL Kinase Buffer provided 90 uL 10X Heparin 1 ug mL 10 pL Your enzyme fraction 10 pL 10 pL 10 pL CK2 Positive Control 2 m unit pL i PTPL Buffer 10 uL 10X Heparin 1 ug mL See page 4 section Materials Required but not Provided Cat CY E1170 1 or Cat CY E1170 2 See page 4 section Materials Required but not Provided 1 Following the above table add the Reagents to each well of the Microplate Finally initiate the reaction by adding 10 uL of
4. Kinase Buffer as needed All samples should be assayed in duplicate W To assay partially purified recombinant CK2 add 10 uL of each fraction to the wells of the assay plate on ice Duplicate wells containing 20 m units 10 uL CK2 Cat CY E1170 1 or Cat CY E1170 2 shouldbe ineluded in each assay as a positive control for phosphorylation 4 Begin the kinase reaction by addition of 90 uL Kinase Reaction buffer per well cover with plate sealer and incubate at 30 C for 30 minutes 5 Wash wells five times with Wash Buffer making sure each well is filled completely Remove residual WashBuffer by gentle tapping or aspiration 6 Pipette 100 uL of HRP conjugated Detection Antibody into each well cover with a plate sealer and incubate at room temperature ca 25 C for 30 minutes Discard any unused conjugate Cat CY 1170 6 Version 141117 SR CK2 KinaseAssay Inhibitor Screening Kit f yeLex User s Manual For Research Use Only Not for use in diagnostic procedures 7 Wash wells five times as same as in step 5 8 Add 100 uL of Substrate Reagent to each well and incubate at room temperature ca 25 C for 5 15 minutes 9 Add 100 uL of Stop Solution to each well in the same order as the previously added Substrate Reagent 10 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 be used Readiithe plate at 450 nm if
5. User s Manual For Research Use Only Not for use in diagnostic procedures Related Products CK2 alpha beta Positive control Cat CY E1170 1 CK2 alpha beta Positive control Cat CY E1170 2 Anti phospho p53 S46 monoclonal antibody TK 4D4 CY M1022 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 http www cyclex co jp CycLex CircuLex products are supplied for research us components thereof may not be resold modified for res products without prior written approval from CycLe such commercial use please contact us via Q C x CircuLex products and ed to manufacture commercial To inquire about licensing for C CY 1170 14 Version 141117
6. Cat CY 1170 10 Version 141117 ry CK2 KinaseAssay Inhibitor Screening Kit ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Example of Test Results Fig 1 Dose dependency of recombinant CK2 enzyme reaction 2 5 p 2 0 F 5 w 7 1 0 F CKIIo CKIIB 0 5 amp CKTla CKTIB 0 0 0 10 20 30 40 50 60 70 80 Enzyme mU reaction Fig 2 Time course of recombinant CK2 enzyme rea 2 5 gt 2 0 F 1 5 p R yt lt 1 0 e CKIla CKIIp 0 5 a amp CKIIa CKIIpB 0 0 1 1 1 1 1 1 0 20 40 60 80 100 120 Reaction time min C CY 1170 11 Version 141117 LA CK2 KinaseAssay Inhibitor Screening Kit Ce ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Fig 3 1 Effect of TBB Calbiochem Cat No 218697 on activity of recombinant CK2 110 100 90 80 IC50 CKII a CKII g 20uM CKII q CKII g 20uM 70 60 CKIlo CKIIp A CKIla CKIIB 50 40 30 F Relative intensity of control 20 0 10 20 30 40 50 60 70 80 90 100 TBB uM Fig 3 2 Effect of Heparin on activity of recombina A 110 1C50 100 CKII q CKII 8 65ng ml 90 CKI a CKII g 20 ng ml S 80 fari 5 g OT e CKila CKIIB S 60 a CKIla CKIIB gt g 5 x g 30 20
7. Your enzyme fraction or Buffer to each well and mixing thoroughly at room temperature Cover with plate sealer Incubate at 30 C for 30 minutes 2 Follow the Standard Assay steps 5 10 page 6 7 Evaluation of Results 1 Average the absorbance values for the CK2ample duplicates positive control and all experimental sample duplicate values when applicable When the CK2 positive control 20 m units assay is included as an internal control for the phosphorylation reaction the absorbance value should be greater than 1 0 with a background less than 0 15 2 For screening of purification chroMatography fractions on graph paper plot the mean absorbance values for each of the samples onjthe Y axis versus the fraction number on the X axis to determine the location of the eluted purified CK2 3 For kinetic analysis on graph paper plot the mean absorbance values for each of the time points on the Y axis versus the time of eal reaction minutes on the X axis Assay Characteristies The CycLex Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit has been shown to detect the activity 0 CK2 in column fractions of human or animal cell lysates The assay shows good linearity of sample response The assay may be used to follow the purification of CK2 Cat CY 1170 9 Version 141117 oy CK2 KinaseAssay Inhibitor Screening Kit P ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Trou
8. bleshooting 1 The CK2 positive control should be run in duplicate 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 of the assay is of the first order Variations in the protocol can lead to non linearity of the curve as can assay kinetics that are other than first order For a non linear curve point to point or quadratic curve fit methods should be used 3 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 allow the plate to dry out Add S bstrat Reagent immediately after wash Reagent Stability All of the reagents included in the CycLex Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit have been tested for stability Reagents shouldsnot be used beyond the stated expiration date Upon receipt kit reagents should be stored at 4 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
9. e CK2 in mammary gland tumorigenesis Oncogene 20 3247 3257 a Seldin D C and Leder P 1995 CK2 alpha transgene induce murine lymphoma relation to theileriosis in cattle Science 267 894 897 oo Landesman Bollag E Channavajhala P L Cardiff RD and Seldin D C 1998 p53 deficiency and misexpression of protein kinase CK2a collaborate in the development of thymic lymphomas in mice Oncogene 16 2965 2974 9 Channavajhala P and Seldin D C 2002 yFun fional interaction of protein kinase CK2 and c Myc in lymphomagenesis Oncogene 21 5280 5288 10 Sayed M Pelech S Wong C Marotta A and Salh B 2001 Protein kinase CK2 is involved in G2 arrest and apoptosis following spindle damage in epithelial cells Oncogene 20 6994 7005 11 Desagher S Osen Sand Ay Montessuit S Magnenat E Vilbois F Hochmann A Journot L Antonsson B and Martinou J C 2001 Phosphorylation of bid by casein kinases I and II regulates its cleavage by caspase 8 Mol Cell 8 601 611 12 Li P Li J Mullen Otto A Dietz R and von Harsdorf R 2002 Phosphorylation by protein kinase CK2 A signaling switch for the caspase inhibiting protein ARC Mol Cell 10 247 258 13 WangaH Davis A Yu S and Ahmed K 2001 Response of cancer cells to molecular interruption of the CK2 signal Mol Cell Biochem 227 167 174 Cat CY 1170 13 Version 141117 Pae CK2 KinaseAssay Inhibitor Screening Kit Ce yCLex
10. ecommendations Special considerations when screening activators or inhibitors In order to estimate the inhibitory effect on CK2 activity in the test chemicals Correctly it is necessary to conduct the control experiment of Solvent control at least once for_evefysexperiment 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 CK2 actiyity 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 3 Assay reagents Test sample Solvent control Inhibitor control Kinase Reaction Buffer 80 uL 80 uL 80 uL 10X Inhibitor or equivalent 10 pL Solvent for Inhibitor 10 pL 10X Heparin 1 pg mL 10 pL CK2 Positive Control 2 m unit uL 10 pL 10 uL 10 uL or your enzyme fraction 10X Heparin 1 ug mL See page 4 section Materials Required but not Provided Cat CY E1170 1 or Cat CY E1170 2 See page 4 section Materials Required but not Provided 1 Following the above table add the Reagents to each well of the microplate Finally initiate reaction by adding 10 uL of Dilut d K2 positive control to each well and mixing thoroughly at room temperature Cover with plate sealer Incubate at 30 C for 30 minutes 2 Follow the Standard Assay steps 5 10 page 6
11. enzyme should be stored in aliquots at 70 C e 10X Heparin 1 wg mL Heparin Na is available from Sigma Cat H 4784 100 pg mL stock solution H20 diluted 1 100 in KinasesBuffer e Pipettors 2 20 uL 20 200 uL and 200 1000 pL precision pipettors with disposable tips Precision repeating pipettor e Wash bottle or multichannel dispenser for plate washing e Microcentrifuge and tubes 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 e 500 or 1000 mL graduated ylinder e Reagent reservoirs Deionized watenof the highest quality e Disposable paper towels Cat CY 1170 4 Version 141117 oe CK2 KinaseAssay Inhibitor Screening Kit aa ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Precautions and Recommendations e 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
12. gat of TK 4D4 an anti phospho p53 serine46 specific antibody which then catalyzes the conversion of h chromogenic substrate tetra methylbenzidine TMB from a colorless solution to a blue solution orjyellow after the addition of stopping reagent The color is quantified by spectrophotometry andffeflects the relative amount of CK2 activity in the sample For kinetic analysis the sample containing K2 is added to the wells in a similar fashion and at varying times the reaction is stopped by the addition of a chelator sodium ethylenediaminetetraacetate EDTA and the amount of phosphorylated substrate determined as before The CycLex Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit is designed to accurately determine the presence and relative amount of CK2 activity in purification column fractions and to determine non isotopic kinetic analysis of CK2 actiwity Careful attention to methods of chromatography and the assay protocol will provide the investigator With a reliable tool for the evaluation of CK2 activity Summary of Procedure Add 100 uL of reaction mix tothe wells 4 Incubate for 30 min at 30 C Wash the wells t Add 100 uL of ARP conjugated anti phosphorylated form specific antibody 4 Incubate for 30 min at room temp Was the wells t Add 100ML of Substrate Reagent t Add 100 uL of Stop Solution t Measure absorbance at 450 nm Cat CY 1170 3 Version 141117 oe CK2 KinaseAssay Inhibitor Screen
13. ing Kit aa 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 afforl zip lock bag with a desiccant pack Wells are coated with recombinant p53 N terminus 1 99 a a as substrate of CK2 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 One vial of lyophilized ATP Naz salt HRP conjugated Detection Antibody One vial containing 12 mL of ARP horseradish peroxidase conjugated anti phospho p53 S46 monoclonal antibody TK 4D4 Ready to lise Substrate Reagent One bottle containing 20 mL of the chromoge nie substrate tetra methylbenzidine TMB Ready to use Stop Solution One bottle containing 20 mL of 1 N H2S04 Ready to tse Materials Required but not Provided e CK2 alpha beta and CK2 alpha beta positive control Available from CycLex Cat CY E1170 1 and Cat CY E1170 2 One vial contains 4 units 100 uL CK2 enzyme Positive control should be added to the first well at 20 m units well For instance diluted positive control 1 20 use 10 uL for 1 assay Unused CK2
14. l and possible implication in viral infections Thi makes CK2 an attractive target for anti neoplastic and antiviral drugs Experimental studies suggest that dysregulated expression of the alpha subunit 6f CK2 imparts an oncogenic potential in the cells such that in cooperation with certain oncogenes it produces a profound enhancement of the tumor phenotype Recent studies have provided evidence that overexpression of CK2 in tumor cells is not simply a reflection of tumor cell proliferation alone but additionally may reflect the pathobiological characteristics of the tumor Of e nsiderable interest is the possibility that CK2 dysregulation in tumors may influence the apoptoti jactivity in those cells 12 Approaches to interfering with the CK2 signal may provide a useful means for inducing tumor cell death 13 Measurement of CK2 activity The protocol generally regarded as most sensitive for the quantitative measurement of CK2 activity involves incubation of the CK2 sample with substrate either a natural or synthetic polypeptide such as CK2 substrate peptide RRRDDDSDDD in the pres ice of Mg and P labeled ATP The reaction is terminated by spotting a sample onto a phosphocellulose P81 filter paper disc followed by washing extensively to remove unincorporated radiolabel and the incorporated radioactivity on P81 filter is counted While sensitive this method is labor infensive generates hazardous radioactive waste and de
15. on of 90 uL 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 incubate at 30 C for 20 minutes 5 Stop the rea tion by fli king out the contents Alternatively the reaction may be terminated by the addition of 150 uE OMFM Na EDTA pH 8 0 to each well 6 Wash wells five 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 HRP conjugated Detection Antibody into each well cover with a plate sealer andincubate at room temperature ca 25 C for 30 minutes Discard any unused conjugate Cat CY 1170 7 Version 141117 CK2 KinaseAssay Inhibitor Screening Kit User s Manual For Research Use Only Not for use in diagnostic procedures wy yclex 8 Wash wells five times as same as in step 6 9 Add 100 uL of Substrate Reagent to each well and incubate at room temperature ca 25 C for 10 15 minutes 10 add 100 uL of Stop Solution to each well in the same order as the previously add d Substrate Reagent 11 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 be used Read the plate at 450 nm if only a single wavelength can be used Wells must be read within 30 mintites of adding the Stop Solution R
16. only a single wavelength can be used Wells must be read within 30 minutes f 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 aspirating or decanting myertithe plate and blot it against clean paper towels Note 2 Reliable signals are obtained when either O D values do not exceed 0 25 units for the blank no enzyme control or 2 5 units for the CK2 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 CK2 activity ofgoff seale samples The readings at 405 nm should not replace the on scale readings at 450 nm Kinetic Assay 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 N Prepare all samples diluted with Kinase Buffer as needed All samples should be assayed in duplicate w To assay partially purified recombinant CK2 add 10 uL of each fraction to the wells of the assay plate on ice Duplicate wells contaiming 20 m units 10 uL CK2 Cat CY E1170 1 or Cat CY E1170 2 should be included in each assay as a positive control for phosphorylation 4 Begin kinase reaction by additi
17. pends on a radioisotope of short half life It s particularly unsuitable when kinase assays are only performed on an infrequent basis The Cyel x Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit uses a peroxidase coupled anti phospho p53 serine46 monoclonal antibody as a reporter molecule in a 96 well ELISA format This assay provides a non isotopic sensitive and specific method to detect CK2 activity Cat CY 1170 2 Version 141117 oe CK2 KinaseAssay Inhibitor Screening Kit cA ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Principle of the Assay The CycLex Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit is a single site semi quantitative immunoassay for CK2 activity Plates are pre coated with a substrate corresponding to recombinant p53 which contains a serine residue that are phosphorylated by CK2 Casein kinase II The detector antibody specifically detects only the phosphorylated form of serine46 on p53 The CycLex Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit can be used to study the kinetics of a purified or partially purified CK2 as well as to screening these kinases inhibitor To perform the test the sample is diluted in Kinase Buffer pipetted into the wells and allowed to phosphorylate the bound substrate in the presence of Mg and ATP The amount of phosphorylated substrate is measured by binding it with a horseradish peroxidase conju
18. rips required for the particular determination Since experimental conditions may vary an aliquot of the CK2 Cat CY E1170 1 or CY E1170 2 available separately from CycLex should be meluded in each assay as a positive control Disposable pipette tips and reagent troughs should be used for all liquid 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 ddH20 to the viakof 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 C 3 Prepare Kinase Reaction Buffer by mixing following reagents 96 assays 10 assays 1 assay Kinase Buffer provided 9 5 mL 950 pL 95 uL 20X ATP Solution 0 5 mL 50 uL 5 uL Total 10 mL 1000 uL 100 uL 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 unused wells t the foi pouch refold seal with tape and store at 4 C N Prepare all samples diluted with
19. ry CK2 KinaseAssay Inhibitor Screening Kit cA ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Non Radioisotopic Kit for Measuring CK2 activity CycLex CK2 Kinase Assay Inhibitor Screening Kit Cat CY 1170 Intended Use ccccccccccccceeceeeeeceeeeeeees 1 Be sat nsatsinanigayonmnysaneiocsiciassadsaepiaaeeeisinen 1 Tntroduction cceceeeecceecseeesssseesessesenseees 2 Principle Of the Assay 3 Materials Provided ceccceeeseeeeeeeeeeees 4 Materials Required but not Provided 4 Precautions and Recommendation 5 Detailed Protocol ccceecceeesseseseeeeeees 6 9 Evaluation of Results cccccceeeeeeeereeee 9 Assay Characteristics reese 9 Troubleshooting s sscasneosssesacesonsnanensnanbncnunsiss 10 Reagent SEADILY sstssnsosacavnandnneaopiamnsonanngndytn 10 Example of Test Results cece 11612 RePCLEN CES ecscdcisssissssasssingiesiebadsiaaceastecssenevae 13 Related Products ccceececeeeseeseeeeeees 14 Intended Use The CycLex Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit designed to measure the activities of purified CK2 fof the rapid and sensitive evaluation of inhibitors or activators The phospho serine specific monoclonal antibody used in this assay kit has been demonstrated to recognize the phospho serine46 residue in p53 which is phosphorylated by CK2 in vitro Applications of this kit include 1
20. the highest quality e Do not mix reagents from different kits The buffers and reagents in this kit may contain preservatives or othemchemi als Care should be taken to avoid direct contact with these reagents e Do not mouth pipet or ingest any of the reagents e Do not smoke eat or drink when performing the assay or in argas where samples or reagents are handled e Dispose of tetra methylbenzidine TMB containifg 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 Solutiongandth 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 CAUTION Sulfuric Acid is a strong acid Wear disposable gloves and eye protection when handling Stop Solution Cat CY 1170 5 Version 141117 oe CK2 KinaseAssay Inhibitor Screening Kit aa ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Detailed Protocol The CycLex Research Product CycLex CK2 Kinase Assay Inhibitor Screening Kit is provided wath removable strips of wells so the assay can be carried out on separate occasions using only the number of st

CY-1170

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