Enzymes
Products & Ordering
| Protein Kinases | |
| RafRBD (c-Raf) PR-305 Ras Binding Domain of Raf, c-Raf 1 Kinase human, recombinant, E. coli | RafRBDGST (C-Raf) PR-366 Ras Binding Domain of Raf, C-Raf 1 Kinase human, recombinant, E. coli |
| Akt1/ PKBαHis – inactive PR-325 Protein Kinase B human, recombinant, Sf9 insect cells | MEK1GST – active PR-161 Mitogen-activated kinase kinase 1, MAPKK1 human, recombinant, E. coli |
| Phosphatases | |
| Cdc25A PR-348 Cell division cycle phosphatase human, recombinant, E. coli | PTP 1B PR-931 Protein Tyrosine Phosphatase 1B human, recombinant, E. coli |
| SHP-1GST – full length PR-947 Src homology 2 domain Phosphatase-1 human, recombinant, E. coli | VHR PR-350 VH1-related phosphatase human, recombinant, E. coli |
| Peptidases | |
| Legumain PR-967 Asparaginyl Endopeptidase (AEP) human, recombinant, Leishmania | Furin PR-968 Proprotein Convertase Subtilisin/Kexin Type 3 (PCSK3) human, recombinant, human cell line HEK293S |
| PC7 PR-969 Proprotein Convertase Subtilisin/Kexin Type 7 (PCSK7) human, recombinant, human cell line HEK293S |
Restriction Enzymes: General Information and Quality Control
Unit Definition
One unit of restriction endonuclease activity is defined as the amount of enzyme required to produce a complete digest of 1 µg of substrate DNA (or fragments) in a total reaction volume of 50 µl in 60 minutes under optimal assay conditions as stated for each restriction endonuclease.Determination of the volume activity of restriction endonucleases
Restriction endonuclease activity assays are performed by adding different enzyme dilutions to the appropriate assay buffer containing 1 µg of substrate DNA. After a 60-minutes incubation at the appropriate temperature, the digestion is stopped and the DNA samples are visualized by agarose gel/ethidium bromide electrophoresis. The most diluted enzyme solution giving a complete digest is used to calculate the activity in units/µl.
Please note that the activity is substrate dependent and when working with a new substrate, the enzyme should be titrated to determine the actual or expected activity.Quality Controls
The results of all quality control assays are reported on the Technical Data Sheet provided with each enzyme.Overdigestion Assay
An overdigestion assay was used as a qualitative determination of enzyme purity and of a lack of nonspecific DNases. In the overdigestion assay, increasing amounts of each restriction endonuclease (usually 10, 20, 30, 40, 50 units) are added to a series of tubes containing 1 µg substrate DNA. After a 20-hours incubation under the recommended assay conditions, the maximum number of units giving a clear, sharp, normal banding pattern is determined by agarose gel/ethidium bromide electrophoresis.
To pass the test, the enzyme must yield an unaltered banding pattern under conditions of up to 600-fold overdigestion (units x hours) as compared to a 2-fold digest. If enzyme exhibits “star” activity at a lower than 600-fold functional excess, the product description includes information on the functional excess at which the “star” activity does not occur.Assay for Nonspecific Endonucleases
To assay for nonspecific endonuclease contamination, each restriction endonuclease is incubated with a supercoiled plasmid substrate lacking the recognition sequence of the restriction endonuclease. A single nonspecific nick in the RF I DNA converts it to the RF II form (nicked circle). Increasing amounts of enzyme (usually 10, 20, 30, 40, 50 units) are added to a series of tubes containing 1 µg of RF I (supercoiled form) DNA. After a 20-hours incubation under the recommended assay conditions, the two forms are distinguished on agarose gels and the percent conversion from RF I to RF II is determined.Ligation and Recutting Assay
A ligation assay was used to determine the functional purity of the DNA after restriction enzyme digestion. Substrate DNA is completely digested with a 10- and 50-fold excess of the restriction endonuclease in the appropriate assay buffer, ligated with T4 DNA Ligase and recut with the same restriction enzyme. Cut, ligated and recut DNAs are analyzed by agarose gel/ethidium bromide electrophoresis. A normal banding pattern indicates intact 5 and 3 termini as well as the absence of contaminating nucleases or phosphatases.Stability
All Jena Bioscience restriction endonucleases are reassayed every 4-6 months. This process allows us to ensure full enzyme activity and optimal performance in every enzyme we ship. Due to the excellent results of this testing, we have extended the expiration dates of most of our enzymes to 18 months.
One unit of restriction endonuclease activity is defined as the amount of enzyme required to produce a complete digest of 1 µg of substrate DNA (or fragments) in a total reaction volume of 50 µl in 60 minutes under optimal assay conditions as stated for each restriction endonuclease.Determination of the volume activity of restriction endonucleases
Restriction endonuclease activity assays are performed by adding different enzyme dilutions to the appropriate assay buffer containing 1 µg of substrate DNA. After a 60-minutes incubation at the appropriate temperature, the digestion is stopped and the DNA samples are visualized by agarose gel/ethidium bromide electrophoresis. The most diluted enzyme solution giving a complete digest is used to calculate the activity in units/µl.
Please note that the activity is substrate dependent and when working with a new substrate, the enzyme should be titrated to determine the actual or expected activity.Quality Controls
The results of all quality control assays are reported on the Technical Data Sheet provided with each enzyme.Overdigestion Assay
An overdigestion assay was used as a qualitative determination of enzyme purity and of a lack of nonspecific DNases. In the overdigestion assay, increasing amounts of each restriction endonuclease (usually 10, 20, 30, 40, 50 units) are added to a series of tubes containing 1 µg substrate DNA. After a 20-hours incubation under the recommended assay conditions, the maximum number of units giving a clear, sharp, normal banding pattern is determined by agarose gel/ethidium bromide electrophoresis.
To pass the test, the enzyme must yield an unaltered banding pattern under conditions of up to 600-fold overdigestion (units x hours) as compared to a 2-fold digest. If enzyme exhibits “star” activity at a lower than 600-fold functional excess, the product description includes information on the functional excess at which the “star” activity does not occur.Assay for Nonspecific Endonucleases
To assay for nonspecific endonuclease contamination, each restriction endonuclease is incubated with a supercoiled plasmid substrate lacking the recognition sequence of the restriction endonuclease. A single nonspecific nick in the RF I DNA converts it to the RF II form (nicked circle). Increasing amounts of enzyme (usually 10, 20, 30, 40, 50 units) are added to a series of tubes containing 1 µg of RF I (supercoiled form) DNA. After a 20-hours incubation under the recommended assay conditions, the two forms are distinguished on agarose gels and the percent conversion from RF I to RF II is determined.Ligation and Recutting Assay
A ligation assay was used to determine the functional purity of the DNA after restriction enzyme digestion. Substrate DNA is completely digested with a 10- and 50-fold excess of the restriction endonuclease in the appropriate assay buffer, ligated with T4 DNA Ligase and recut with the same restriction enzyme. Cut, ligated and recut DNAs are analyzed by agarose gel/ethidium bromide electrophoresis. A normal banding pattern indicates intact 5 and 3 termini as well as the absence of contaminating nucleases or phosphatases.Stability
All Jena Bioscience restriction endonucleases are reassayed every 4-6 months. This process allows us to ensure full enzyme activity and optimal performance in every enzyme we ship. Due to the excellent results of this testing, we have extended the expiration dates of most of our enzymes to 18 months.
Restriction Enzymes: "A" Enzymes
| Products & Ordering | |
| AccI EN-E2004 Isoschizomers: FblI, XmiI 5’–GT↓MK AC–3’ 3’–CA KM↑TG–5′ | AluI EN-101 JBSpeed Restriction Enzyme 5’–AG↓CT–3’ 3’–TC↑GA–5’ |
| ApaI EN-E2015 Neoschizomers: Bsp120I, PspOMI JBSpeed Restriction Enzyme 5’–G GGCC↓C–3’ 3’–C↑CCGG G–5′ | ApaLI EN-172 Isoschizomers: Alw44I JBSpeed Restriction Enzyme 5’–G↓TGCA C–3’ 3’–C ACGT↑G–5’ |
| AsuII EN-102 Isoschizomers: Bpu14I, Bsp119I, BspT104I, BstBI, Csp45I, NspVI, SfuI JBSpeed Restriction Enzyme 5’–TT↓CG AA–3’ 3’–AA GC↑TT–5′ | AvaI EN-E2006 Isoschizomers: Ama87I, AvaI, BsiHKCI, BsoBI, Eco88I, NspIII Neoschizomers: BmeT110 I 5’–C↓YCGR G–3’ 3’–G RGCY↑C–5’ |
| AvaII EN-E2007 Isoschizomers: AflI, Bme18I, Eco47I, SinI, VpaK11BI JBSpeed Restriction Enzyme 5’–G↓GWC C–3’ 3’–C CWG↑G–5′ | Universal Restriction Buffer EN-300 10 x conc. |
Restriction Enzymes: "B" Enzymes
| Products & Ordering | ||
| BamHI EN-103 JBSpeed Restriction Enzyme 5’–G↓GATC C–3’ 3’–C CTAG↑G–5′ | BanII EN-E2060 Isoschizomers: Eco24I, EcoT38I, FriOI JBSpeed Restriction Enzyme 5’–G RGCY↓C–3’ 3’–C↑YCGR G–5′ | BclI EN-104 Isoschizomers: FbaI, Ksp22I JBSpeed Restriction Enzyme 5’–T↓GATC A–3’ 3’–A CTAG↑T–5’ |
| BglI EN-105 5’–GCCN NNN↓NGGC–3’ 3’–GCCN↑NNN NGGC–5′ | BglII EN-106 5’–A↓GATC T–3’ 3’–T CTAG↑A–5′ | BseAI EN-107 Isoschizomers: AccIII, Bsp13I, BspEI, Kpn2I, MroI JBSpeed Restriction Enzyme 5’–T↓CCGG A–3’ 3’–A GGCC↑T–5’ |
| BseBI EN-108 Isoschizomers: BstNI, BstOI, Bst2UI, MvaI, Neoschizomers: AjnI, EcoRII, Psp6I, PspGI JBSpeed Restriction Enzyme 5’–CC↓W GG–3’ 3’–GG W↑CC–5′ | BseCI EN-109 Isoschizomers: BanIII, Bsa29I, Bsp106I, BspDI, BspXI, Bsu15I, BsuTUI, ClaI, ZhoI JBSpeed Restriction Enzyme 5’–AT↓CG AT–3’ 3’–TA GC↑TA–5′ | BshFI EN-110 Isoschizomers: BspANI, BsuRI, HaeIII, PalI, PhoI JBSpeed Restriction Enzyme 5’–GG↓CC–3’ 3’–CC↑GG–5’ |
| BsiSI EN-111 Isoschizomers: HapII, HpaII, MspI JBSpeed Restriction Enzyme 5’–C↓CG G–3’ 3’–G GC↑C–5′ | BssAI EN-112 Isoschizomers: Bse118I, BsrFI, Cfr10I JBSpeed Restriction Enzyme 5’–R↓CCGG Y–3’ 3’–Y GGCC↑R–5′ | BssHII EN-E2115 Isoschizomers: BsePI, PauI 5’–G↓CGCG C –3’ 3’–C GCGC↑G–5’ |
| BstEII EN-144 Isoschizomers: BstPI, Eco91I, EcoO65I, PspEI 5’–G↓GTNAC C–3’ 3’–C CANTG↑G–5′ | BstXI EN-E2118 JBSpeed Restriction Enzyme 5’–CCAN NNNN↓NTGG–3’ 3’–GGTN↑NNNN NACC–5′ | Universal Restriction Buffer EN-300 10 x conc. |
Restriction Enzymes: "C" Enzymes
Restriction Enzymes: "D" Enzymes
Restriction Enzymes: "E" Enzymes
Restriction Enzymes: "F" Enzymes
Restriction Enzymes: "H" Enzymes
| Products & Ordering | ||
| HincII EN-E2200 Isoschizomers: HindII JBSpeed Restriction Enzyme5’–GTY↓RAC–3’3’–CAR↑YTG–5′ | HindIII EN-116 JBSpeed Restriction Enzyme5’–A↓AGCT T–3’3’–T TCGA↑A–5′ | HinfI EN-117 JBSpeed Restriction Enzyme5’–G↓ANT C–3’3’–C TNA↑G–5’ |
| HpaI EN-118 Isoschizomers: KspAI JBSpeed Restriction Enzyme5’–GTT↓AAC–3’3’–CAA↑TTG–5′ | HpaII EN-E2260 Isoschizomers: BsiSI, HapI, MspI5’–C↓CG G–3’3’–G GC↑C–5′ | Universal Restriction Buffer EN-300 10 x conc |
Restriction Enzymes: "K" Enzymes
Restriction Enzymes: "M" Enzymes
| Products & Ordering | ||
| MboI EN-120 Isoschizomers: BfuCI, Bsp143I, BstENII, BstMBI, DpnII, Kzo9II, NdeII Neoschizomers: BstKTI JBSpeed Restriction Enzyme5’–↓GATC –3’3’– CTAG↑–5′ | MboII EN-E2284 JBSpeed Restriction Enzyme5’–GAAGANNNNNNN N↓–3’3’–CTTCTNNNNNNN↑N –5′ | MluI EN-E2287 JBSpeed Restriction Enzyme5’–A↓CGCG T–3’3’–T GCGC↑A–5’ |
| MnlI EN-E2289 JBSpeed Restriction Enzyme5’–CCTCNNNNNN N↓–3’3’–GGAGNNNNNN↑N –5′ | MspCI EN-121 Isoschizomers: AflII, BfrI, BspTI, Bst98I, Vha464I JBSpeed Restriction Enzyme5’–C↓TTAA G–3’3’–G AATT↑C–5′ | Universal Restriction Buffer EN-300 10 x conc |
Restriction Enzymes: "N" Enzymes
| Products & Ordering | |
| NaeI EN-122 Isoschizomers: PdiI, Neoschizomers: MroNI, NgoMIV5’–GCC↓GGC–3’3’–CGG↑CCG–5′ | NarI EN-E2291 Isoschizomers: Mly113I, Neoschizomers: BbeI, DinI, EgeI, EheI, KasI, SfoI5’–GG↓CG CC–3’3’–CC GC↑GG–5’ |
| NcoI EN-123 Isoschizomers: Bsp19I JBSpeed Restriction Enzyme5’–C↓CATG G–3’3’–G GTAC↑C–5′ | NdeI EN-E2293 Isoschizomers: FauNDI JBSpeed Restriction Enzyme5’–CA↓TA TG–3’3’–GT AT↑AC–5’ |
| NheI EN-146 Isoschizomers: AsuNHI, Neoschizomers: BmtI JBSpeed Restriction Enzyme5’–G↓CTAG C–3’3’–C GATC↑G–5′ | NotI EN-124 Isoschizomers: CciNI5’–GC↓GGCC GC–3’3’–CG CCGG↑CG–5’ |
| NruI EN-125 Isoschizomers: Bsp68I5’–TCG↓CGA–3’3’–AGC↑GCT–5′ | Universal Restriction Buffer EN-300 10 x conc. |
Restriction Enzymes: "P" Enzymes
| Products & Ordering |
| PspPI EN-126 Isoschizomers: AspS9I, Cfr13I, Sau96I JBSpeed Restriction Enzyme5’–G↓GNC C–3’3’–C CNG↑G–5’ |
| PstI EN-127 Isoschizomers: BspMAI5’–C TGCA↓G–3’3’–G↑ACGT C–5’ |
| PvuI EN-E2320 Isoschizomers: BpvUI, BspCI, MvrI, Ple19I JBSpeed Restriction Enzyme5’–CG AT↓CG–3’3’–GC↑TA GC–5’ |
| PvuII EN-128 JBSpeed Restriction Enzyme5’–CAG↓CTG–3’3’–GTC↑GAC–5’ |
| Universal Restriction Buffer EN-300 10 x conc. |
Restriction Enzymes: "R" Enzymes
Restriction Enzymes: "S" Enzymes
| Products & Ordering | ||
| SalI EN-130 5’–G↓TCGA C–3’3’–C AGCT↑G–5′ | ScaI EN-131 Isoschizomers: Acc113I, ZrmI JBSpeed Restriction Enzyme5’–AGT↓ACT–3’3’–TCA↑TGA–5′ | SfiI EN-132 5’–GGCCN NNN↓NGGCC–3’3’–CCGGN↑NNN NCCGG–5’ |
| SgrBI EN-133 Isoschizomers: Cfr42I, KspI, SacII, Sfr303I5’–CC GC↓GG–3’3’–GG↑CG CC–5′ | SlaI EN-134 Isoschizomers: BssHI, PaeR7I, Sfr274I, TliI, XhoI5’–C↓TCGA G–3’3’–G AGCT↑C–5′ | SmaI EN-135 Neoschizomers: Cfr9I, PspAI, XmaI, XmaCI5’–CCC↓GGG–3’3’–GGG↑CCC–5’ |
| SnaBI EN-136 Isoschizomers: BstSNI, Eco105I JBSpeed Restriction Enzyme5’–TAC↓GTA–3’3’–ATG↑CAT–5′ | SpeI EN-E2398 Isoschizomers: AhlI, BcuI JBSpeed Restriction Enzyme5’–A↓CTAG T–3’3’–T GATC↑A–5′ | SphI EN-137 Isoschizomers: BbuI, PaeI, SpaHI5’–G CATG↓C–3’3’–C↑GTAC G–5’ |
| SseBI EN-138 Isoschizomers: AatI, Eco147I, PceI, StuI5’–AGG↓CCT–3’3’–TCC↑GGA–5′ | SspI EN-139 JBSpeed Restriction Enzyme5’–AAT↓ATT–3’3’–TTA↑TAA–5′ | SstI EN-140 Isoschizomers: Psp124BI, SacI, Neoschizomers: Ecl136II, EcoICRI5’–G AGCT↓C–3’3’–C↑TCGA G–5’ |
| StyI EN-141 Isoschizomers: BssT1I, Eco130I, EcoT14I, ErhI5’–C↓CWWG G–3’3’–G GWWC↑C–5′ | Universal Restriction Buffer EN-300 10 x conc. |
Restriction Enzymes: "T" Enzymes
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