Data Sheet
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1. er vial 1 ml of 10X assay buffer mixed with 9 4 C Quantity required 25 uL assay Dilute the 50X Fluoro Substrate Pepti ith 1 1X Assay buffer 3 2X Fluoro Deacetylated Peptide 4 uoro Deacetylated Peptide Quantity required 25 uL assay Dilute the 3 50X Fluoro Deac ted Peptide 1 25 with 1 1X Assay buffer 4 X20 diluted Lysylendpep Quantity required 5 uL ass U ml 20 with 1 1X Assay buffer 5 HDAC stop Quantity Requir ml Lysylendpeptidase and 2 uM Trichostatin A e Mix following 00 uL 4 assays Lysylendpeptidase 2 pL 2 200X Trichostatin A 2 uL 3 1 1X Assay buffer 196 uL C CY 1150 5 Versions 130130 Kc HDACs Deacetylase Fluorometric Assay Kit es Mey User s Manual For Research Use Only Not for use in diagnostic procedures 6 One step assay buffer Final 0 25 mAU ml Lysylendpeptidase and 20 uM Fluoro Substrate Pep in 50 uL of assay mixture Quantity Required 30 uL assay in case of adding 10 uL of enzyme and 10 uL of inhibito equivalent Mix following reagents 30 uL 1 assay 1 DIOX Assay buffer 5yuL 2 50X Fluoro Substrate Peptide l uL 3 4 X20 diluted Lysylendpeptidase 2 5uL 4 Distilled water 21 5 uL 7 5X Inhibitor or equivalent 5X final concentration of Quantity required 10 uL assay Dilute Inhibitor or equivalent to 5X final desired concentration wi2. 12 Version 130130 P HDACs Deacetylase Fluorometric Assay Kit f ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Fig 3 Effect of Trichostatin A on HDAC activity One step method Trichostatin A 100nM Trichostatin A 10nM 4 Trichostatin A 1nM DMSO md un o S o o 2 X Q Ke S uw Ve en i 40 60 80 HDAC reaction time min HDA C substrate CycLex HDAC Assay kit 10 000 70 000 E 60 000 S 50 000 E a crude HDAC amp 40 000 e ja 30 000 e Recombinant 30 000 SIRTI E di en E 0 10 20 30 40 50 60 Time min C CY 1150 13 Version 130130 P HDACs Deacetylase Fluorometric Assay Kit f ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Fig 5 Substrate Preference of HDAC and SIRT1 using CycLex Sir2 kit lt Sir2 substrate CycLex Sir2 Assay kit gt e I0 000 255555550 cuu ee S 9 000 5 8 000 4 7 000 2 6 000 crude HDAC LC 5 000 S 4 000 Recombinant 3 000 SIRT1 IC 2 000 E 1 000 g E TEM 0 LL op odo dod od odo dod d dod d d d d 1 1 1 d 1 1 1 1 di 1 1 0 0 10 20 30 40 50 60 Time min Fig 6 Time course of 2 reaction in a Two step meth NW reaction 400 350 300 r 5 S 250 T 200 E e No enzyme control amp 150 nA vi e No in
3. agents on HDACs Crude HDAC at 70 C and all other components below 20 C gents to excessive light use only and not for use in diagnostic or therapeutic procedures CY 1150 Versions 130130 4 HDACs Deacetylase Fluorometric Assay Kit vyclex User s Manual For Research Use Only Not for use in diagnostic procedures Introduction Histone deacetylase HDAC is considered to play a crucial role in regulating gene expressio changing nucleosome structure HDAC is also thought to participate in regulation of cell cy differentiation and it has been reported that the failure of this regulation leads to some types Inhibition of HDAC activity by HDAC inhibitors such as trichostatin A TSA and s hydroxamic acid SAHA induce differentiation and or apoptosis of transformed cells in v tumor growth in a mouse model It has been reported that HDAC inhibitors are effective fo treatment of acute promyelocytic leukemia APL and various cancers Thus HDAC inhibitors are expected to function as new anti tumor drugs and antibacterial reagents It is thoug at screening of histone deacetylase inhibitors is likely to be further carried out as one way to r additional substances with similar properties However the conventional method for measuring HDAC activity is very c and laborious In order to measure HDAC enzyme activity it is necessary to prepare radioact lated histone as a substrate First cells have to be labeled metabolicall
4. HDACs Deacetylase Fluorometric Assay Kit d e Mey User s Manual For Research Use Only Not for use in diagnostic procedures Quantitative test kit for histone deacetylase activity CycLex HDACs Deacetylase Fluorometr Assay Kit ays Cat CY 1150 Intended E 1 Qy Geet 1 UE 2 Principle of The Assay eerie 3 Materials Provided iswcccscssvsanccanserevasecacceseansas 3 Materials Required but not Provided 4 PRECAUTIONS rn r Detailed PROG eiscissstnorsevadcsrsessvicguatensiuanes CC PIOUS SN c Troubleshootin iusso oreet Reagent Stability ees tees cem irren Sample Preparation eeeeeee Example of Test Results Referenc s ee Ee Related Droducts eee Intended Use The CycLex Research Product CycLe activity in lysates Primarily the CycLe X acetylase Fluorometric Assay Kit detects HDAC esearch Product CycLex HDACs Deacetylase Fluorometric Assay Kit is designed for the rapid and sensitive evaluation of HDAC inhibitors using crude HDAC fraction Additionally any cultured ell cell line or tissue homogenate can be assayed for HDAC activity with the CycLex e roduct CycLex HDACSs Deacetylase Fluorometric Assay Kit if the appropriate dose of HDA cific inhibitor e g Tricostatine A is used Applications for this kit incl 1 Monitoring the purificatio ACs including HDACI 2 3 and 8 2 Screening inhibitors of activators of HDACs 3 Detecting the As acological
5. P Anti Human SIRT1 Rabbit Polyclonal Antibody Cat CY P1016 NAD Dependent Deacetylase SIRT1 Cat CY E1151 NAD Dependent Deacetylase SIRT2 Cat CY E1152 NAD Dependent Deacetylase SIRT3 Cat CY E1153 NAMPT Nicotinamide Phosphoribosyltransferase Cat CY E125 NMNATI Nicotinamide Mononucleotide Adenylyltransferase E1252 Note This product is covered under CycLex s patents U S Patent No 7 033 778 and No 7256013 European Patent No 1243658 Japanese Patent No 4267043 Canadian Patent No 2392711 1063 103 Terasawaoka Ina Nagano 396 0002 Japan Fax 81 265 76 7618 e mail info cyclex cog PRODUCED BY CycLex Co Ltd e s are supplied for research use only CycLex CircuLex products and y not be resold modified for resale or used to manufacture commercial ior written approval from CycLex Co Ltd To inquire about licensing for please contact us via email components t products wit CY 1150 Versions 130130
6. action G 2 Incubate for 20 min or desired length of time at room temperature 3 Add 50 uL of 5 HDAC stop solution to each well 4 Incubate for at least 10 min or desired length of time at made between 10 minutes and 40 minutes perature Measurement should be 5 Read fluorescence intensity using microtiter pla me ith excitation at 355 nm and emission at 460 nm 6 During the time in which HDAC reaction rate is maintained the difference in fluorescence intensity between No inhibitor control and No G or inhibitor control indicates the HDAC activity Note 1 Although the above table indicatesthe yolume of addition of 7 5X Inhibitor or equivalent as 10 uL the concentration a e me of test reagents to add can be changed so that the concentration of test rea es the setting concentration For example since the final volume of reaction is 50 is also possible to add 5 uL of 10X test reagent In this case please add 1 1X A ffer to set to the final reaction volume of 50 uL Note 2 Although the vol be changed to a v buffer to set Note 3 ole tion of Your enzyme sample is set to 10 uL in this table it may e up to 20 uL at your discretion In that case please add 1 1X Assay action volume of 50 uL nt is recommended C CY 1150 8 Version 130130 wy HDACs Deacetylase Fluorometric Assay Kit L Vcl ex User s Manual For Research Use Only Not for use in diagnos
7. der plates Considering that the use of fully automatic apparatus to measure fluorescence become widespread HDAC activity measurement which could not be made by the conven is now possible with the CycLex HDACs Deacetylase Fluorometric Assay Kit using the same equipment This new method of measurement should dramatically raise the efficiency of inhibiter screening and biochemical analysis of these enzymes Measuring Principle of The CycLex HDACs Deacetylase Fluoro X X X Lys Ac MCA 4 Deacetylase X X X Lys MCA v Lysly endpepti X X X Lys AMC Measurement of fluoresce e Note This measuring principle and kit are covere der CycLex s patents U S Patent No 7 033 778 and No 7256013 European Patent No 1243658 Japanese Patent No 4267043 Q Canadian Patent No 2392711 Materials Provided Each kit contains CA 1 1 9 e Iml x2 Below 20 C Peptide 1 mM acetylated Peptide 1 mM 50uLx1 Below 20 C Lysylendpeftidase 100 mAU ml S0uLx1 Below 20 C CrudeffD XC crude nuclear extract from HeLa D Wstructionmanual LL room temp CY 1150 Versions 130130 HDACs Deacetylase Fluorometric Assay Kit d e Mey User s Manual For Research Use Only Not for use in diagnostic procedures Materials Required but not Provided Microplate for fluorometer Microplate reading fluorometer capable of excitation at a wavelength i
8. ev 9 40 84 1999 3 Fenrick R amp Hiebert S W J Cell Biochem Suppl 30 31 194 202 1998 4 Yoshida M Horinouchi S amp Beppu T Bioassays 17 423 430 1995 Q 5 Richon V M et al Proc Natl Acad Sci USA 93 5705 5708 1996 A 6 Richon V M et al Proc Natl Acad Sci USA 95 3003 3007 1998 Q 7 Cohen L et al Proc AACR 39 108 abstr 736 1998 8 Desai D El Bayoumy K amp Amin S Proc AACR 40 2396 abs 1999 9 Laherty C D Yang W M et al Cell 89 349 356 1997 11 Hoffmann K Grosch G amp Jung M Nucleic Acids Res 27 2057 2058 1999 C CY 1150 16 Version 130130 P HDACs Deacetylase Fluorometric Assay Kit t ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Related Products CycLex Cellular Histone Acetylation Assay Kit Cat CY 1140 CycLex HDACs Deacetylase Fluorometric Assay Kit Cat CY 1150 CycLex HDACS Deacetylase Fluorometric Assay Kit Cat CY 1158 CycLex SIRT1 Sir2 Deacetylase Fluorometric Assay Kit Cat CY 1151 CycLex SIRT2 Deacetylase Fluorometric Assay Kit Cat CY 1152 CycLex SIRT3 Deacetylase Fluorometric Assay Kit Cat CY 1153 CycLex SIRT6 Deacetylase Fluorometric Assay Kit Cat CY 1156 Anti Acetylated Histone p53 K382 Mouse Monoclonal Antibody Cat CY M10 Anti Histone Deacetylase 1 HDAC1 Rabbit Polyclonal Antibody Cat CY P Anti Histone Deacetylase 2 HDAC2 Rabbit Polyclonal Antibody Cat CY
9. glycerol Procedure Isolation of Nuclei Suspend 1x 10 cells 100 mm dish sub conflu into 1ml of lysis buffer Vortex for 10 second Keep on ice for 15 min Spin the cells through 4 ml of sucrose at 1 300 x g for 10 min at 4 C Discard the supernatant Wash the nuclei pellet once with c 0 Tris HCl pH7 5 10 mM NaCl Ei Un opa R3 rs Extraction of Nuclei Suspend the isolated nuclei i uL of extraction buffer Sonic ate for 30 seconds Stand on ice for 30 min c f g at 20 000 x g for Take supernatant t Clear extract by Bradford method or equivalent tract at 70 C until use Determine protein kind of protease inhibitor DEN Ee po ci Store the crude Ki Note C CY 1150 11 Version 130130 P HDACs Deacetylase Fluorometric Assay Kit c Mey User s Manual For Research Use Only Not for use in diagnostic procedures Example of Test Results Fig 1 Dose dependency of crude HDAC Two step method 500 450 400 350 300 250 200 Dd 150 100 50 Relative crude HDAC activity x10 counts 0 2 4 6 8 10 Crude HDAC Dose ul NJ Fig 2 Time course of HDAC reaction Two step met 600 Un e 4L e G No enzyme control groom ITE c peeing No inhibitor control k Inhibitor control w e t2 e F355 F460 x10 counts eme EI e S 0 20 40 60 80 HDAC reaction time min C CY 1150
10. have an inhibitory effect on lysylendpeptidase are mixed in a fraction purified from various cells or the immunoprecipitate using a specific antibody a or other proteins precise HDAC enzyme activity cannot be measured Since the protease mhibitors used in the usual protein purification process inhibit lysylendpeptidase activity str please avoid the use of any protease inhibitors during the protein purification process 3 Final fluorescence intensity will not increase both when test chemicals hay HDAC and also when there is an inhibitory effect on lysylendpeptidase itory effect on 360 380 nm and cannot be evaluated correctly 5 The Crude HDAC should be run in duplicate using the proto d in the Detailed Protocol Incubation times or temperatures significantly different fr ecified may give erroneous 4 If the test reagents themselves emit fluorescence at excitation fluorescence wavelength 440 460 nm the inhibitory effect of the t results 6 The reaction curve is nearly a straight line if the ki he assay is of the first order Variations in the protocol can lead to non linearity of the curv n kinetics that are other than first order For a non linear curve point to point or quadra fit methods should be used instructions in the Detailed Protocol were 7 Poor duplicates indicate inaccurate dispensing If followed accurately such results indicate a for multi channel pipettor maintenance Reagent Stabilit
11. hibitor control E 100 4 Inhibitor control 50 Q 0 10 20 30 40 Reaction Time min C CY 1150 14 Version 130130 HDACs Deacetylase Fluorometric Assay Kit es Mey User s Manual For Research Use Only Not for use in diagnostic procedures Fig 7 Measurement of HeLa cell endogenous HDACI in an immunoprecipitate using anti HD antibody Cat CY P1011 by means of CycLex HDACs Deacetylase Fluorometric Assay Kit e Anti HDAC 1 pAb e 6 000 000 e Normal rabbit IgG 5 000 000 4 000 000 3 000 000 2 000 000 Fluorescence Intensity F355 F460 1 000 000 0 3 10 15 20 25 30 Reaction Time min Fig 8 Measurement of HeLa cell endogenous antibody Cat CY P1012 by means of Cyc an immunoprecipitate using anti HDAC2 ACs Deacetylase Fluorometric Assay Kit e Anti HDAC 2 pAb 6 000 000 6 Normal rabbit IgG S 5 000 000 o E Ke e T 4 000 000 gt E 3 000 000 8 5 o 2 000 000 S A E 1 000 000 0 OO09OO0009000090090900009 0 5 10 15 20 25 30 Reaction Time min C CY 1150 15 Versions 130130 Kc HDACs Deacetylase Fluorometric Assay Kit es Mey User s Manual For Research Use Only Not for use in diagnostic procedures References Davie J R amp Chadee D N J Cell Biochem Suppl 30 31 203 213 1998 2 Kouzarides T Curr Opin Genet D
12. method and the 2 steps method In the the reaction is initiated and the fluorescence intensity is measured by mixing fluorescence labeled acetylated peptide which is substrate HDAC and lysly endpeptida i reaction is not stopped it is necessary to measure fluorescence intensity at regular intervals after the reaction is initiated and to determine reaction velocity Alternatively within a time i hich the reaction velocity is kept constant it is also possible to stop the reaction by adding tricho 4 an HDAC inhibitor and to measure fluorescence intensity Conversely the 2 step method b initiating a reaction of fluorescence labeled acetylated peptide and HDAC within a set ti 10d to remove an acetyl group from substrate peptide then in the second step adds lysly endpept trichostatin A as an HDAC enzyme inhibitor to stop the HDAC reaction while simultan ving the resultant deacetylated fluorescence labeled peptide by lysly endpeptidase Preparation Method for Assay Reagents Thaw 250X Fluoro Substrate Peptide and 3 50X Fluoro Deacety eptide at room temperature Stand other reagents in ice to thaw Use them after they thaw co tely 1 1X Assay buffer 20 mM Tris HCl pH 8 0 125 mM l g Quantity Required 100 uL assay Dilute the 10X Assay buffer 1 10 with disti Since this is the base buffer for the assay pre ml distilled water and store 10 ml of assay buffe 2 2X Fluoro Substrate Peptide 40 uM C Peptide ycerol
13. n the range 350 380 an detection of emitted light in the range 440 460 nm Pipettors 2 20 uL 20 200 uL and 200 1000 uL precision pipettors with disposable ti multi channel pipette Microplate shaker Deionized water of the highest quality 500 or 1000 mL graduated cylinder Reagent reservoirs Precautions Peptide at room are completely thawed here is a possibility that the enzyme activity may be inactivated Aliquot to 10 20 uL and 0 C e Please avoid mixing of protease inhibitors such as PMSF measured HDAC activity e in the sample that will be Do not use kit components beyond the indicated ki Rinse all detergent residue from glassware Use deionized water of the highest quality Do not mix reagents from different kits Do not mouth pipette or ingest any of t nts e Do not smoke eat or drink whe ng the assay or in areas where samples or reagents are handled Biological samples may be wounds or breathe aeros inated with infectious agents Do not ingest expose to open protective gloves and dispose of biological samples properly C CY 1150 4 Versions 130130 P HDACs Deacetylase Fluorometric Assay Kit f ycLex User s Manual For Research Use Only Not for use in diagnostic procedures Detailed Protocol Description of assay system CycLex HDACs Deacetylase Fluorometric Assay Kit can measure the enzyme activi with two kinds of measuring methods the 1 step
14. tein purification If there is such a possibility please carry out the of Assay control using Fluoro Deacetylated Peptide to reference When Fluoro eptide is used fluorescence intensity should increase whenever there is no HDAC activi a sample When there is an inhibitory effect on lysylendpeptidase activity even if there is should not increase 4 Not only when an inhibitory effect on HDAC is i chemicals but also when there is an inhibitory effect on lysylendpeptidase final fluorescence Mintensity will not increase Please use Fluoro Deacetylated Peptide instead of Fluore Substrate Peptide and conduct a control experiment that does not add HDAC Although fluores tensity increases even if HDAC is not added when Fluoro Deacetylated Peptide is used when itory effect on lysly endpeptidase activity occurs in a test sample fluorescence intensity d ot ease For research use only not for use in dia c or therapeutic procedures Ki d Ki d C CY 1150 9 Version 130130 HDACs Deacetylase Fluorometric Assay Kit 4 Vclex User s Manual For Research Use Only Not for use in diagnostic procedures Troubleshooting Although trichostatin A is added in the 2 steps method in order to stop a HDAC reaction the acti of Sir2 and it s human homologue SIRT1 cannot be measured correctly even if NAD is added sinc they do not have susceptibility in trichostatin A Please use CycLex s Sir2 assay kit 2 When chemicals that
15. th 1 1X ffer 8 5X Trichostatin A 5 uM Trichostatin A Quantity Required 10 uL assay Dilute the 5 200X Trichostatin A 1 40 with 1 1X Assay buffer 9 X10 diluted crude HDACS 1 10 diluted crude nuclear extract fr eLa Quantity required 10 uL assay Dilute the Crude HDAC 1 10 with 1 1X Assay buffer Note Use 9 X10 diluted crude HDACs within the same day they are prepared C CY 1150 6 Version 130130 yelex Assay Procedures 1 One step method User s Manual HDACs Deacetylase Fluorometric Assay Kit For Research Use Only Not for use in diagnostic procedures or Your enzyme sample No enzyme No inhibitor Assay reagents Test sample control control 6 One step assay buffer 30 uL 30 uL 30 uL 1 1X Assay buffer 20 uL 10 uL 7 5X Inhibitor or equivalent 10 uL 8 5X TSA 10 uL 9 X10 diluted crude HDACs 10 uL 10 pL 10 pL 1 Following the above table add Reagent 6 1 7 and 8 to each initiate reaction by adding 10 uL of 9 X10 diluted crude HDACs and mixing thoroughly at room temperature 2 Read fluorescence intensity for 30 to 60 minutes at to 2 fluorometer with excitation at 355 nm and emission at 460 reaction while the reaction velocity remains constant Alternate procedure 1 Following the above table add Reagent 6 initiate reaction by adding 10 uL of 9 X10 di and mixing thoroughly at room tempera
16. tic procedures Cautions 1 In order to measure the activity of HDAC correctly it is necessary to conduct the control experi for No enzyme control and Inhibitor control at least once in addition to No inhibitor cont indicated in the above table Although fluorescence intensity increases in No inhibite when HDAC enzyme activity is in the sample the increase in fluorescence intensity is No enzyme control and Inhibitor control 2 In order to estimate the inhibitory effect on HDAC activity in the test chemicals correetly it is necessary to conduct the control experiment of No inhibitor control at least once for every experiment and Inhibitor control at least once for the first experiment in additi est sample as indicated in the above table When test chemicals cause an inhibitory effect on activity the level of increase of fluorescence intensity is weakened as compared with i control The increase in fluorescence intensity is not observed in Inhibitor control e to be mixed in crude the specific antibody st be measured Since the y inhibit lysylendpeptidase 3 When the chemicals that have an inhibitory effect on lysylendpeptid HDAC fraction purified from various cells or the immunoprecipita against HDAC or other proteins precise HDAC enzyme activi protease inhibitors used in the usual protein purification proce activity please avoid using any protease inhibitors during the p o
17. ture 2 While the reaction rate is kept constant a to stop the reaction and measure fluoresce excitation at a wavelength in the range nm 3 The difference in fluorescence i it indicates the HDAC activity d C CY 1150 2 d 8 to each well of the microplate Finally rude HDACs or your enzyme to each well C rvals using microtiter plate ure and calculate the rate of microplate Finally enzyme to each well L of 8 5X TSA to each well at appropriate time t nsity in a microplate fluorescence reader capable of and detection of emitted light in the range 440 460 etween No inhibitor control and No enzyme control Version 4 HDACs Deacetylase Fluorometric Assay Kit t Mey User s Manual For Research Use Only Not for use in diagnostic procedures 2 Two step method Test No enzyme No inhibitor Inhibitor Assay reagents sample control control control t 1 1X Assay buffer 5 pL 25 uL 15 pL 5 L 2 2X Fluoro Substrate Peptide 25 uL 25 uL 25 uL 2 7 5X Inhibitor or equivalent 10 pL 8 5X TSA 10 3 2X Fluoro Deacetylated Peptide 25 uL 9 X10 diluted crude HDACs 10 uL 10 pL L or Your enzyme sample Your enzyme sample 10 uL 1 Following the table above add Reagent 1 2 7 and 3 to each we X10 diluted crude HDACs or your enzyme to each well and mix tho add 10 uL of 9 to initiate re
18. y All of the reagents included in the C search Product HDAC Assay Kit have been tested for stability Reagents should not be u the stated expiration date Upon receipt store the 6 Crude HDAC at 70 C all other ea s should be stored below 20 C Ki E C CY 1150 10 Version 130130 s HDACs Deacetylase Fluorometric Assay Kit t Vclex User s Manual For Research Use Only Not for use in diagnostic procedures Sample Preparation Numerous extraction and purification methods can be used to isolate HDACs The follo protocols have been shown to work with a number of different cells and enzyme sources provided as examples of suitable methods Crude samples can frequently be used without diluti more concentrated or highly purified HDACs should be diluted It is strongly advised always perform an initial experiment to determine the proper dilution to be used experiments This need not be any more than a single time point assay using serial dilution S Ould be extract cell lysate or sample fraction taken prior to a purification step All sample preparatio performed at 4 C and recovered fractions should be kept at 70 C to prevent loss of e atic activity Buffers J Lysis Buffer Sucrose cushion Extraction buffer 10 mM Tris HCl pH7 5 30 Sucrose 50 mM Hepes KOH pH 10 mM NaCl 10 mM Tris HCl pH7 5 7 5 15 mM MgCl 10 mM NaCl 420 mM NaCl 250 mM Sucrose 3 mM MgCl 0 5 mM EDTA Na 0 5 NP 40 0 1 mM EGTA 0 1 mM EGTA 10
19. y with radioactivity b Wg radioactive acetic acid to the culture medium Second radioactive acetylated histone has be purified from the cells Following the reaction it is necessary to extract and separate the radio been released from acetylated histone using ethyl acetate to measure the radioactivity Although a method for measuring the activity of deacetylase wi se of radioactive substances was reported in recent years owing to the use of fluorescent labeled a ated lysine as a substrate the reaction product must be separated from the intact substrate fluorescent intensity measured by reverse phase HPLC As mentioned above these measure stems are difficult to adapt for processing many samples under a variety of conditio their complicated operation Thus a simple system for biochemical analysis as well as reening without the use of radioactive substances is preferred y of the enzyme based on C CY 1150 2 Versions 130130 P HDACs Deacetylase Fluorometric Assay Kit vcl ex User s Manual For Research Use Only Not for use in diagnostic procedures Principle of the Assay CycLex HDACs Deacetylase Fluorometric Assay Kit measures the activity of HDAC by the E principle of changing an HDAC reaction into the activity of the protease Since it is very simple measure common protease activity and it can be performed at a low price the measuremen activity in most laboratories is possible if they are equipped with a fluorescent rea
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