Home
Data Sheet - MBL Life science
Contents
1. U Poor duplicates accompanied by elevatedivalues 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 notiallow the plate to dry out Add Substrate Reagent immediately after wash Reagent Stability All of the reagents included in the CycLex Research Product CycLex Pim 1 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 CY 1167 11 Version 120420 7 yclex Example of Test Results For Research Use Only Not for use in diagnostic procedures Pim 1 Kinase Assay Inhibitor Screening Kit User s Manual Fig 1 Dose dependency of recombinant Pim 1 enzyme reaction 3 5 3 0 5 10 15 20 Pim 1 Positive control m units Fig 2 Time course of recombinant Pim 1 enzyme r action Cat CY 1167 A450 2 5 2 0 1 5 1 0 0 5 0 0 10
2. Japan Fax 81 265 76 7618 e mail info eyclex co jp URL http 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 CY 1167 14 Version 120420
3. ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Evaluation of Results 1 Average the absorbance values for the Pim 1 sample duplicates positive control and all experimental sample duplicate values when applicable When Pim 1 positive control 10 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 kinetic analysis on graph paper plot the mean absorbance values for each of the time points on the Y axis versus the time of each reaction minutes on the X axis Assay Characteristics The CycLex Research Product CycLex Pim 1 Kinase Assay Inhibitor Screening Kit has been shown to detect the activity of purified recombinant Pim 1 The assay shows good linearity of sample response Troubleshooting The CycLex Pim 1 positive control Cat CY E1167 should be r n 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 of the assay is of the first order Variations in the protocol can lead to non linearity of the curve 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
4. sensitive and specific methodto measure the activities of Pim 1 Cat CY 1167 2 Version 120420 P Pim 1 Kinase Assay Inhibitor Screening Kit ey cA ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Principle of the Assay The CycLex Research Product CycLex Pim 1 Kinase Assay Inhibitor Screening Kit is a single site semi quantitative immunoassay for Pim 1 activity Plates are pre coated with a substrate corresponding to recombinant p21wafl which contains threonine residues that can be efficiently phosphorylated by Pim 1 The detector antibody specifically detects only the phosphorylated form of threonine145 residue on p2lwafl The CycLex Pim 1 Kinase Assay Inhibitor Screening Kit may be usedet0 study the kinetics of a purified Pim 1 as well as to screening Pim 1 inhibitor or activator To perform thestest the sample is diluted in Kinase Buffer pipetted into the wells and allowed to phosphorylatesthe bound substrate following the addition of Mg and ATP The amount of phosphorylated substrate is measured by binding it with a PWT 01 a anti phospho p21wafl threonine145 polyclonal antibody followed by binding with horseradish peroxidase conjugated anti rabbit 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
5. the relative amount of Pim 1 activity in the sample For kinetic analysis the Pim 1 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 Vv Incubate for 30min at 30 C Wash the wells Add 100 uL of antiephospho threonine polyclonal antibody PWT 01 Vv Incubate for 30 min at room temp Wash the wells Add 100 uL of HRP conjugated anti rabbit IgG y Incubate for 30 min at room temp Wash the wells v Add 100 uL of Substrate Reagent v Add 100 pL of Stop Solution v Measure absorbance at 450 nm Cat CY 1167 3 Version 120420 Pim 1 Kinase Assay Inhibitor Screening Kit yy 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 p2l1wafl as Pim 1 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 f
6. 20 30 40 50 60 Reaction Time min 12 Version 120420 A Pim 1 Kinase Assay Inhibitor Screening Kit ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Fig 3 Km for ATP recombinant Pim 1 y 9 9676x 671 58 R 0 9805 2000 nN oS oS So So A gt Ts n V R N r A 50 100 150 ATP conc uM lt s gt Fig 4 Effect of broad spectrum kinase inhibitor staurosporine and K252a on Pim 1 kinase activity 110 p Staurosporine e K252a p e Oo 90 80 70 60 50 40 30 20 10 0 ems ac 0 0001 0 001 0 01 0 1 1 10 Drug conc uM Rerated intensity of control Cat CY 1167 13 Version 120420 pry Pim 1 Kinase Assay Inhibitor Screening Kit ycLex User s Manual For Research Use Only Not for use in diagnostic procedures References 1 Cuypers H T Selten G Quint W Zijlstra M Maandag E R Boelens W van Wezenbeek P Melief C and Berns A 1984 Cell 37 141 150 2 Selten G Cuypers H T and Berns A 1985 EMBO J 4 1793 1798 3 Selten G Cuypers H T Boelens W Robanus Maandag E Verbeek J Domen J van Beveren C and Berns A 1986 Cell 46 603 611 4 Nagarajan L Louie E Tsujimoto Y Ar Rushdi A Heubner K and Croce C M 1986 Proc Natl Acad Sci U S A 83 2556 2560 5 Amson R Sigaux F Prezedborski S Flan
7. 1 Applications of this kit include 1 Screening inhibitors or activators of Pim 1 2 Detecting the effects of pharmacological agents on Pim 1 activity This assay kit is for research use only and not for use in diagnostic or therapeutic procedures Storage Upon receipt store all components at 4 C e Don t xpose reagents to excessive light Cat CY 1167 1 Version 120420 P Pim 1 Kinase Assay Inhibitor Screening Kit oy cdl ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Introduction The pim 1 oncogene encodes a serine threonine kinase Pim 1 involved in the transduction of cytokine triggered mitogenic signals The pim 1 oncogene was originally identified as a genetic locus frequently activated by the proviral insertion of Moloney murine leukemia virus into mouse T cell lymphomas 1 3 The pim 1 oncogene has also been implicated in human hematopoietic malignancies with its overexpression frequently detected in human hematopoietic cell lines as well as in fresh tumor cells from patients with leukemia 4 5 During embryonal development the pim 1 gene is expressed mainly in developing fetal hematopoietic tissues 5 The pim 1 gene product Pim 1 identified as a serine threonine kinase 6 8 has been thought to play a critical role in the transduction of mitogenic signals from cytokines since Pim 1 expression is rapidly induced after cytokine stimulation and the pr
8. addition to Test sample as indicated in the following table When test chemicals cause an inhibitory effect on Pim 1 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 3 or Purified enzyme sample Assay reagents Test sample Solvent Inhibitor control control Kinase Reaction buffer 80 uL 80 pL 80 uL 10X Inhibitor or equivalent 10 uL Solvent for Inhibitor 10 pL 10X Staurosporine 20 uM 10 uL Pim 1 positive control 1 m unit pL 10 uL 10 uL 10 uL Cati S 4400 See Page 4 section Materials Required but not Provided Pim 1 positive control Cat CY E1167 See Page 5 section Materials Required but not Provided Cat CY 1167 9 Version 120420 yy wyclex User s Manual Pim 1 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 Pim 1 positive control to each well and mixing thoroughly at room temperature Cover with plate sealer or lid and incubate at 30 C for 30 minutes 2 Follow the Standard Assay steps 5 12 page 8 Special considerations when measuring precise Pim 1 kinase activity In order to measure the activity of Pim 1 correctly it is necessary to conduct th
9. al antibody PWT 01 into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 30 minutes 7 Wash wells five times as same as in step 5 8 Pipette 100 uL of HRP conjugated anti rabbit IgG into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 30 minutes Discard any unus d 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 atoom 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 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 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 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 25 units for the blank no enzyme control or 2 5 units forthe Pim 1 positive control Note 3 If the microplate reader is not capable of reading absorbance greater than the absorbance of the Weel positive control pe
10. drin G Givol D and Telerman A 1989 Proc Natl Acad Sci U S A 86 8857 8861 Saris C J M Domen J and Berns A 1991 EMBO J 10 655 664 7 Padma R and Nagarajan L 1991 Cancer Res 51 2486 2489 8 Hoover D Friedmann M Reeves R and Magnuson N S 1991 J Biol Chem 266 14018 14023 9 Miura O Miura Y Nakamura N Quelle F W Witthuhn B A sThle J N and Aoki N 1994 Blood 84 4135 4141 10 Yip Schneider M T Horie M and Broxmeyer H E 1995 Blood 85 3494 3502 11 O Farrell A M Ichihara M Mui A L F and Miyajima A 1996 Blood 87 3655 3668 12 van der Lugt N M T J Domen E Verhoeven K Linders H yvan der Gulden J Allen and A Berns 1995 EMBO J 14 2536 13 van Lohuizen M S Verbeek P Krimpenfort J Domen C Saris T Radaszkiewicz and A Berns 1989 Cell 56 673 14 Acton D J Domen H Jacobs M Vlaar S Korsmeyer and A Berns 1992 Curr Top Microbiol Immunol 182 293 15 Moroy T A Grzeschiczek S Petzold and K WU Hartmann 1993 Proc Natl Acad Sci USA 90 10734 16 Nosaka T T Kawashima K Misawa kK Ikuta A L F Mui and T Kitamura 1999 EMBO J 18 4754 17 Shirogane T T Fukada J M M Muller D T Shima M Hibi and T Hirano 1999 Immunity 11 709 an Related Products Pim 1 Positive control Gat CY E1167 PRODUCED BY CycLex Co Ltd 1063 103 Terasawaoka Ina Nagano 396 0002
11. e 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 of A450 increases in Test sample when Pim 1 enzyme activity is in the sample thehighlevel 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 20 uM 10 uL Purified enzyme sample 10 uL 10 uL 10 uL Pim 1 positive control 1 m unit uL 10 uL Buffer 10 uL Cat S 4400 See Page 4 section Materials Required but not Provided Pim 1 positive control Cat CY E1167 See Page 5 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 Purified enzym sample or Buffer to each well and mixing thoroughly at room temperature Cover with plate sealer or lid and incubate at 30 C for 30 minutes 2 Follow the Standard Assay steps 5 12 page 8 Cat CY 1167 10 Version 120420 pry Pim 1 Kinase Assay Inhibitor Screening Kit
12. hich 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 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 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 1167 6 Version 120420 yy YCLEX Detailed Protocol User s Manual Pim 1 Kinase Assay Inhibito
13. illed completely Remove residual Wash Buffer by gentle tapping or aspiration 7 Pipette 100 uL of anti phospho p21wafl threonine145 polyclonal antibody PWT 0 into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 30 minutes 8 Wash wells five times as same as in step 6 9 Pipette 100 uL of HRP conjugated anti rabbit IgG into each well cover with plate sealer or lid and incubate at room temperature ca 25 C for 30 minutes Discard any unused conjugate after use 10 Wash wells five times as same as in step 6 11 Add 100 uL of Substrate Reagent to each well and incub te at room temperature ca 25 C 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 spectrophotometric plate reader at dual wavelengths of 450 540 nm Dual wavelengths of 450 550 or450 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 Pim 1 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
14. oliferative response to cytokines is impaired in cells from pim 1 deficient mice 9 1 1 When overexpressed pim genes can efficiently cooperate with myc or bel 2 oncogenes in lymphomagenesis 12 14 Pim 1 has been reported to protect thymocytes against glucocorticoid induced apoptosis 15 and to promote cell proliferation or survival in severahIL 3 or IL 6 dependent hematopoietic cell lines 16 17 Measurement of Pim 1 Kinase activity The protocol generally regarded as most sensitive for the quantitative measurement of Pim 1 activity involves incubation of the Pim 1 sample with substrate either a natural or synthetic polypeptide such as histone H1 substrate peptide in the presence of Mg and P labeled ATP The reaction is terminated by spotting a sample onto a filter paper disc followed by immersion in acid to precipitate the radiolabeled product The filter papers are then washed 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 Pim 1 Kinase Assay Inhibitor Screening Kit uses anti phospho p21wafl threonin 145 polyclonal antibody PWT 01 and peroxidase coupled anti rabbit IgG antibody as a reporter molecule in a 96 well ELISA format This assay provides a non isotopic
15. or Kinase Reaction Buffer and sample dilution 20X ATP Lyophilized ATP Naz salt Reconstitute contents of vial with 0 8 mL of ddH O See section Preparation of Working Solution page 7 Anti phospho p21wafl threonine145 polyclonal antibody PWT 01 One vial containing 12 mL of anti phospho p21wafl threonine145 polyclonal antibody PWT 0L Ready to use HRP conjugated anti rabbit IgG One vial containing 12 mL of HRP horseradish peroxidase conjugated anti rabbit IgG Ready to use Substrate Reagent One bottle containing 20 mL of the chromogenic substrate tetra methylbenzidine TMB Ready to use Stop Solution One bottle containing 20 mL of WN H3SO1 Ready to use Cat CY 1167 4 Version 120420 Pim 1 Kinase Assay Inhibitor Screening Kit 4 ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Materials Required but not Provided e Pim 1 positive control Available from CycLex Pim 1 positive control Cat CY E1167 One vial containing 2 units 100 uL Pim 1 enzyme Positive control should be added to the first well at 10 m units well For instance For instance diluted positive control 1 20 with enzyme dilution buffer use 10 uL for 1 assay Unused Pim 1 enzyme should be stored in aliquots at 70 C The Pim 1 Positive control should be diluted with an enzyme dilution buffer to avoid inactivating the enzym vactivity in low protein concentration condition e Enzyme dilution buffer Mi
16. pry Pim 1 Kinase Assay Inhibitor Screening Kit ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Non Radioisotopic Kit for Measuring Pim 1 Kinase Activity CycLex Pim 1 Kinase Assay Inhibitor Screening Kit Cat CY 1167 T tended USS eisicscacsdisdavasinadanslenciaveateexanieniann 1 SAONE ke1 SEE TA 1 T trodu ctiON seners meiit 2 Principle of the Assay esssessessesensesseeeee 3 Materials Provided cccccccccessseceeeesseeeee 4 Materials Required but not Provided 5 Precautions and Recommendation 6 Detailed Protocol ccccccccccccessseeeeeestseeees 7 10 Evaluation of Results ccccsccceeeseeeee 11 Assay Characteristics csccscesscserceeeeees 11 Troubleshooting sxxscsvseseecraHsscasadodnedserewcasl divs 11 Reagent Stability sviiciecsassasiadesaresneiereaeinnictend 11 Example of Test Results ceceeeeeeees 12 13 MPCs gs casctcvhesuktenenesd aan 14 Related Products cs soscstesssststasioveainsentengames 14 Intended Use The CycLex Research Product Cy LexPim 1 Kinase Assay Inhibitor Screening Kit is designed to measure the activities of purified Pim 1 Kinase for the rapid and sensitive evaluation of inhibitors using recombinant Pim 1 The phospho threonine specific polyclonal antibody used in this assay kit has been demonstrated to recognize the phospho threoninel45 residue in p2lwafl which is efficiently phosphorylated by Pim
17. r Screening Kit For Research Use Only Not for use in diagnostic procedures The CycLex Pim 1 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 Pim 1 positive control Cat CY E1167 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 ofthe 20X ATP Solution should be 2 5 mM Store the solution in small aliquots e g 100 uL at 20 O 3 Prepare Kinase Reaction Buffer ATP plus by mixing followmg reagents 96 assays 10 assays 1 assay Kinase Buffer provided 9 5 mL 950 uL 95 uL 20X ATP provided 0 5 mL 50 uL 5 uL Total 10mL 1000 uL 100 pL You will need 80 90 uL of Kinase Reaction Buffer ATP plus per assay well Mix well Discard any unused Kinase Reaction Buffer ATP pl
18. rform a second reading at 405 nm A new O D values measured at 405 nm is used to det tmine Pim 1 activity of off scale samples The readings at 405 nm should not replace the on seale readings at 450 nm Kinetic Assay 1 Remove theappropriateynumber of microtiter wells from the foil pouch and place them into the well holder Returmany 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 34 To assay partially purified recombinant Pim 1 add 10 uL of each fraction to the wells of the assay plate Onpice Duplicate wells containing 10 m units 10 uL of Pim 1 positive control Cat CY E1167 should be included in each assay as a positive control for phosphorylation 4 Begin kinase reaction by addition of 90 uL Kinase Reaction Buffer in duplicate per well in timed intervals suggested interval is 4 minutes but should be individually determined for each system After the final addition incubate at 30 C for 20 minutes Cat CY 1167 8 Version 120420 Pim 1 Kinase Assay Inhibitor Screening Kit 4 gt ycLex User s Manual For Research Use Only Not for use in diagnostic procedures 5 Stop the reaction by flicking out the contents Alternatively the reaction may be terminated bysthe addition of 150 uL 0 1 M Na EDTA pH 8 0 to each well 6 Wash wells five times with Wash Buffer making sure each well is f
19. us after use 4 Prepare Enzyme dilution buffer by mixing 9 parts of Kinase buffer and 1 part of 10X BSA 100 ug mL which is supplied with Pim 1 positive control The Pim 1 Positive control should be diluted with an enzyme dilution buffer to avoid inactivating the enzyme activity in low protein oncentration condition 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 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 To assay partially purified recombinant Pim 1 add 10 uL of each fraction to the wells of the assay plate on ice Duplicate wells containing 10 m units 10 uL of Pim 1 positive control Cat CY E1167 should be included 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 Cat CY 1167 Version 120420 P Pim 1 Kinase Assay Inhibitor Screening Kit oy cA ycLex User s Manual For Research Use Only Not for use in diagnostic procedures 5 Wash wells five 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 p21wafl threonine145 polyclon
20. x 9 parts of Kinase buffer and 1 part of 10X BSA 100 pg mL x 0 25 mL which is supplied with Pim 1 positive control e 10X Staurosporine 20 uM Staurosporine is available from Sigma Cat S 4400 2 mM stock solution DMSO diluted 1 100 in Kinase Buffer e Pipettors 2 20 uL 20 200 uL and 200 1000 uL precision pipettors with disposable tips e 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 an 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 e Reagent reservoirs Deionized water of the highest quality e Disposable paper towels Cat CY 1167 5 Version 120420 Pim 1 Kinase Assay Inhibitor Screening Kit 4 ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Precautions and Recommendations e Store the Pim 1 enzyme at 70 C and the ATP at 20 C when not in use Store all other components at 4 C Do not 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 w
Download Pdf Manuals
Related Search
Related Contents
La Clim, c`est Airwell. presson ville numerique la smart city mode d emploi User Guide - Smart Witness MTX Audio 622CM User's Manual GH3 User manual - Healthcare Lifting Specialists SatGen v3 Manual LevelOne FCS-7111 P4M845 (INTEL i845 Chipset, S-478) Mirco-ATX Form Network Management Card Minislot Electro-Voice P2000 User's Manual Copyright © All rights reserved.
Failed to retrieve file