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A.G. Scientific, Inc, an ISO 9001:2008 company
Phone: (858) 452-9925, Toll Free: (877) 452-9925, Fax: (858) 452-9926
Email: info@agscientific.com, Web site: www.agscientific.com
Certificate of Analysis
PRODUCT: Turbo3C™ (HRV3C) Protease
PRODUCT NUMBER: H-1192
LOT NUMBER: F1269E
DESCRIPTION: Human rhinovirus 3C protease (HRV3C Protease) is a cysteine protease that
recognizes the cleavage site of Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro. It cleaves
between Gln and Gly. The recombinant form of the HRV3C Protease is a
restriction grade protease that has robust activity at 4o
C with high specific
activity and great stability. It does not require any special buffer for its activity
and can be used in a buffer most suitable for target proteins. This HRV3C
Protease is a 47 kDa protein with both GST and His-tags so it can be easy
removal of Turbo3c™ Protease together with cleaved GST- or His-tag in a single
step by either Ni-chelating or Glutathione (GSH) resin.
COMPONENT AND
FORMULATION:
HRV3C Protease: 2 mg/ml in 25 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM
TCEP, and 50% glycerol. Buffer is compatible with Ni-chelating resin and GSH
resin for on-column digestion.
SOURCE: E. coli
SPECIFIC ACTIVITY: The activity of HRV3C Protease is tested using a control target protein. One (1)
µg of HRV3C Protease has at least 1 unit activity conventionally used by other
suppliers (1 unit of HRV3C Protease cleaves >95% of 100 µg of control target
protein at 4o
C for 16 hours).
No non-specific cleavage of the target protein has been observed under the
same condition with HRV3C Protease to control target protein ratio of 1:10
(w/w) of Turbo3c™ Protease to control target protein. Prolonged incubation
(several days) under the same condition does not show any non-specific
cleavage.
A.G. Scientific, Inc, an ISO 9001:2008 company
Phone: (858) 452-9925, Toll Free: (877) 452-9925, Fax: (858) 452-9926
Email: info@agscientific.com, Web site: www.agscientific.com
PROTEASE ACTIVITY:
FIGURE 1
A 68 kDa GST-fusion protein (C) at 1 mg/ml is incubated with HRV3C
Protease (*) at a ratio of (1) 1:50, (2) 1:100, (3) 1:200, (4) 1:400
(w/w) in a buffer of 25mM Tris-HCl, pH 8.0, 150mM NaCl, 14mM β-
mercaptoethanol at 4o
C for 16 hours. The cleaved products are 42 kDa
and 26 kDa.
CLEAVAGE
CONDITION:
It is recommended to use HRV3C Protease at a protease-to-target protein ratio
of 1:100 (w/w) or 1 unit of HRV3C Protease to 100 µg of target protein in a
buffer suitable for the target protein at 4o
C overnight, with the target protein
concentration at 1-2 mg/ml. In most cases, target proteins are completely
cleaved with a protease-to-target protein ratio of 1:50 to 1:400 or 1 unit
HRV3C Protease 50-400 µg of target protein (as shown in Figure 1). The
efficiency of cleavage may vary due to the sequences around the cleavage
site, the conformation and the solubility of the target protein. Due to its high
specificity, more HRV3C Protease (at 1:10 ratio) or longer cleavage time (over
a weekend) at higher temperature (37C) can be used to achieve high cleavage
efficiency without non-specific cleavage of target proteins.
REMOVAL AFTER
CLEAVAGE:
The HRV3C Protease contains both GST and His tags. After cleavage of the
target protein, HRV3C Protease can be easily removed along with the tags from
the cleavage reaction by affinity chromatography on a Ni-chelating resin for
His-tagged target protein or GSH resin for GST-tagged target protein.
STORAGE &
HANDLING:
Store HRV3C Protease at -20o
C. HRV3C Protease is stable at room
temperature for at least two weeks without loss of any activity. It retains full
activity in storage buffer after incubation at 37C for one week.
CAUTION: For laboratory research & scientific manufacturing use only. Not for human or drug use. The
pharmacological and toxicological properties of this product have not been fully investigated. Use caution when
handling. Do not use this compound if you are not fully trained or are unaware of the hazards involved.
Verified: EV

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Hrv 3 c protease

  • 1. A.G. Scientific, Inc, an ISO 9001:2008 company Phone: (858) 452-9925, Toll Free: (877) 452-9925, Fax: (858) 452-9926 Email: info@agscientific.com, Web site: www.agscientific.com Certificate of Analysis PRODUCT: Turbo3C™ (HRV3C) Protease PRODUCT NUMBER: H-1192 LOT NUMBER: F1269E DESCRIPTION: Human rhinovirus 3C protease (HRV3C Protease) is a cysteine protease that recognizes the cleavage site of Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro. It cleaves between Gln and Gly. The recombinant form of the HRV3C Protease is a restriction grade protease that has robust activity at 4o C with high specific activity and great stability. It does not require any special buffer for its activity and can be used in a buffer most suitable for target proteins. This HRV3C Protease is a 47 kDa protein with both GST and His-tags so it can be easy removal of Turbo3c™ Protease together with cleaved GST- or His-tag in a single step by either Ni-chelating or Glutathione (GSH) resin. COMPONENT AND FORMULATION: HRV3C Protease: 2 mg/ml in 25 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM TCEP, and 50% glycerol. Buffer is compatible with Ni-chelating resin and GSH resin for on-column digestion. SOURCE: E. coli SPECIFIC ACTIVITY: The activity of HRV3C Protease is tested using a control target protein. One (1) µg of HRV3C Protease has at least 1 unit activity conventionally used by other suppliers (1 unit of HRV3C Protease cleaves >95% of 100 µg of control target protein at 4o C for 16 hours). No non-specific cleavage of the target protein has been observed under the same condition with HRV3C Protease to control target protein ratio of 1:10 (w/w) of Turbo3c™ Protease to control target protein. Prolonged incubation (several days) under the same condition does not show any non-specific cleavage.
  • 2. A.G. Scientific, Inc, an ISO 9001:2008 company Phone: (858) 452-9925, Toll Free: (877) 452-9925, Fax: (858) 452-9926 Email: info@agscientific.com, Web site: www.agscientific.com PROTEASE ACTIVITY: FIGURE 1 A 68 kDa GST-fusion protein (C) at 1 mg/ml is incubated with HRV3C Protease (*) at a ratio of (1) 1:50, (2) 1:100, (3) 1:200, (4) 1:400 (w/w) in a buffer of 25mM Tris-HCl, pH 8.0, 150mM NaCl, 14mM β- mercaptoethanol at 4o C for 16 hours. The cleaved products are 42 kDa and 26 kDa. CLEAVAGE CONDITION: It is recommended to use HRV3C Protease at a protease-to-target protein ratio of 1:100 (w/w) or 1 unit of HRV3C Protease to 100 µg of target protein in a buffer suitable for the target protein at 4o C overnight, with the target protein concentration at 1-2 mg/ml. In most cases, target proteins are completely cleaved with a protease-to-target protein ratio of 1:50 to 1:400 or 1 unit HRV3C Protease 50-400 µg of target protein (as shown in Figure 1). The efficiency of cleavage may vary due to the sequences around the cleavage site, the conformation and the solubility of the target protein. Due to its high specificity, more HRV3C Protease (at 1:10 ratio) or longer cleavage time (over a weekend) at higher temperature (37C) can be used to achieve high cleavage efficiency without non-specific cleavage of target proteins. REMOVAL AFTER CLEAVAGE: The HRV3C Protease contains both GST and His tags. After cleavage of the target protein, HRV3C Protease can be easily removed along with the tags from the cleavage reaction by affinity chromatography on a Ni-chelating resin for His-tagged target protein or GSH resin for GST-tagged target protein. STORAGE & HANDLING: Store HRV3C Protease at -20o C. HRV3C Protease is stable at room temperature for at least two weeks without loss of any activity. It retains full activity in storage buffer after incubation at 37C for one week. CAUTION: For laboratory research & scientific manufacturing use only. Not for human or drug use. The pharmacological and toxicological properties of this product have not been fully investigated. Use caution when handling. Do not use this compound if you are not fully trained or are unaware of the hazards involved. Verified: EV