Site-Specific Proteases and Cleavage Capture Kits
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| Life Science Research > Protein Sample Preparation > Protein Purification > Kits & Enzymes for Fusion Tags |
Remove fusion tags with restriction-grade site-specific proteases (e.g., Thrombin, Biotinylated Thrombin, Factor Xa, HRV 3C protease, Tag•off™ High Activity recombinant enterokinase [rEK], and rEK) and cleavage capture kits. EKapture™ and Xarrest™ agaroses are used for the removal of rEKs and Factor Xa, respectively, following cleavage of fusion proteins. Biotinylated thrombin is removed with streptavidin agarose and HRV 3C is removed with Ni-NTA agarose.
Features & Benefits
- Highly efficient, specific cleavage of fusion proteins
- Recombinant proteases (such as rEK) have higher activity than native protein
- Highly purified
- Functionally tested for activity, free of detectable contaminating proteases
- Supplied with cleavage control protein and buffers
- Capture kits also available
Applications
Removal of Fusion Tags
Product Performance
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| Description | Components | Unit Definition | Recognition Sequence |
| Thrombin | |||
| Restriction-Grade Thrombin | 50 U Thrombin; 1 mL 10X Thrombin Cleavage Buffer; 2 mL 1X Thrombin Dilution/Storage Buffer; 10 µg Cleavage Control Protein | One unit is defined as the amount of enzyme needed to cleave 1 mg of fusion protein in 16 hours at 20 °C in a 200 µL reaction containing 20 mM Tris-HCl pH 8.4, 150 mM NaCl, 2.5 mM CaCl2, 50 µg fusion protein. | LeuValProArg↓GlySer |
| Biotinylated Thrombin | 50 U Biotinylated Thrombin; 1 mL 10X Thrombin Cleavage Buffer; 2 mL 1X Thrombin Dilution/Storage Buffer; 10 µg Cleavage Control Protein | ||
| Thrombin Cleavage Capture Kit | 50 U Biotinylated Thrombin; 5 x 1 mL 10X Thrombin Cleavage Buffer; 2 mL 1X Thrombin Dilution/Storage Buffer; 2 x 0.4 mL Streptavidin Agarose; 10 µg Cleavage Control Protein; pkg/10 Spin Filters, 2 mL capacity | ||
| Restriction Grade Bovine Factor Xa | |||
| Restriction-Grade Factor Xa | 100 µg Restriction Grade Factor Xa; 2 mL Factor Xa Dilution/Storage Buffer; 1 mL 10X Factor Xa Cleavage Buffer; 10 µg Xa Cleavage Control Protein | One µg of Restriction Grade Factor Xa cleaves 50 µg Xa Cleavage Control Protein to >95% completion in 16 hours at 25 °C in a buffer containing 50 mM Tris-HCl pH 8.0, 100 mM NaCl, and 5 mM CaCl2. | IleGluGlyArg↓ |
| Factor Xa Cleavage Capture Kit | 100 µg Restriction Grade Factor Xa ; 2 mL Factor Xa Dilution/Storage Buffer ; 5 mL 10X Factor Xa Cleavage/Capture Buffer; 2 × 2.5 mL Xarrest™ Agarose; 10 µg Xa Cleavage Control Protein; pkg/10 Spin Filters, 2 mL capacity | ||
| Recombinant Bovine Enterokinase | |||
| Recombinant Enterokinase | 50 U Recombinant Enterokinase; 2 mL 1X rEK Dilution/Storage Buffer; 1 mL 10X rEK Cleavage Buffer; 10 µg Cleavage Control Protein | One unit is defined as the amount of enzyme that will cleave 50 µg of fusion protein in 16 hours at 23 °C, in a buffer containing 20 mM Tris-HCl pH 7.4, 50 mM NaCl, 2 mM CaCl2. | AspAspAspAspLys↓ |
| Enterokinase Cleavage Capture Kit | 50 U Recombinant Enterokinase; 2 mL 1X rEK Dilution/Storage Buffer; 5 mL 10X rEK Cleavage/Capture Buffer; 1.5 mL EKapture™ Agarose ; 10 µg Cleavage Control Protein; pkg/10 Spin Filters, 2 mL capacity | ||
| Tag•off High Activity rEK | 50 U Tag•Off™ High-activity rEK; 10 µg Cleavage Control Protein; 2 mL 1X rEK Dilution/Storage Buffer; 1 mL 10X rEK Cleavage Buffer | One unit of Tag•off™ High Activity rEK is defined as the amount of enzyme needed to cleave 50 µg of fusion protein in 16 hours at 23 °C in a buffer containing 20 mM Tris-HCl, 50 mM NaCl, and 2 mM CaCl2, pH 7.4. | AspAspAspAspLys↓ |
| Tag•off rEK Cleavage Capture Kit | 50 U Tag•off™ High Activity rEK; 10 µg Cleavage Control Protein; 2 mL 1X rEK Dilution/Storage Buffer; 5 mL 10X rEK Cleavage/Capture Buffer; 1.5 mL EKapture™ Agarose; 10 Spin Filter, 2 mL | ||
| HRV 3C Protease | |||
| HRV 3C Protease | 500 U HRV 3C Protease; 10 µg HRV 3C Cleavage Control Protein; 10 mL 10X HRV 3C Cleavage Buffer | One unit of HRV 3C protease is defined as the amount of enzyme that willl cleave >95% of 100 µg His•Tag® fusion control protein in 50 mM Tris-HCl, 150 mM NaCl, pH 7.5 at 4 °C for 16 h. | LeuGluValLeuPheGln↓GlyPro |