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Trypsin - Assay
Source:
Bovine Pancreas
CAS:
9002-07-7
EC:
3.4.21.4
Trypsin Assay
Method
Various assays have been described in the literature. Spencer et al. (1975) report an assay using as substrate an intact protein to which has been attached a fluorescent dye, 1-anilino-8-naphthalenesulfonate (ANS). Stewart (1973) has described a method using chromogenic substrates N-benzoyl-DL-arginine p-nitroanilide (DL-BAPA), N-glutaryl-L-phenylaline p-nitroanilide (L-GPNA). Ford et al. (1973) report on the use of P-nitro-phenyl p-guanidino benzoate (NPGB) to determine active site exposure of immobilized trypsin, a stable acylated enzyme is formed, plus p-nitrophenol spectrophotometrically determined at 410 nm. The assay used in the Worthington laboratory is described below.
One unit hydrolyzes 1 umole of p-toluene-sulfonyl-L-arginine methyl ester (TAME) per minute at 25°C, pH 8.2, in the presence of 0.01 M calcium ion.
Reagents
- 0.046 M Tris⋅HCl buffer, pH 8.1 with 0.0115 M calcium chloride
- 0.01 M TAME (p-toluene-sulfonyl-L-arginine methyl ester)
- 0.001 N HCl
Enzyme
Dilute to a concentration of 10-20 ug/ml in 0.001 N HCl.
Procedure
Set spectrophotometer at 247 nm and 25°C.
Pipette into each cuvette as follows:
0.046 M Tris⋅HCl buffer, pH 8.1 2.6 ml 0.01 M TAME 0.3 ml
Incubate in spectrophotometer at 25°C for 3-4 minutes to achieve temperature equilibration and establish a blank rate, if any. Add 0.1 ml diluted enzyme and record A247 for 3-4 minutes. Determine ΔA247 from initial linear portion of the curve. The reaction remains linear to an A247 of about 0.320. The reaction should be linear for at least three minutes. If this is not so repeat using less enzyme.
Calculation
where 540 is the extinction coefficient of p-toluenesulfonyl-L-arginine at 247 nm
1 TAME Unit = 19.2 USP/NF Units = 57.5 BAEE Units.
Trypsin Products
Description
Activity
Code
Cat. #
Size
Price
Description
SequENZ® Trypsin, Modified, Sequencing Grade
Source:
Bovine Pancreas
Trypsin, treated with L-(tosylamido-2-phenyl) ethyl chloromethyl ketone to inhibit contaminating chymotryptic activity, chemically modified to promote stability and further purified to remove autolysis fragments, resulting in a highly stable trypsin product resistant to autolysis while retaining specificity.
Store at -20°C. PROTECT FROM MOISTURE.
Ice Pack required
Activity
≥150 units per mg protein (at least 8,625 BAEE/2875 USP/NF units per mg protein)
Code
TRSEQZ
Product details
LS02120
4x25 µg
$98.00
LS02122
4x100 µg
$275.00
LS02123
1 mg
$500.00
LS02124
Bulk
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Description
SequENZ® Trypsin, Modified, Sequencing Grade, Solution
Source:
Bovine Pancreas
Ready to use liquid preparation of Trypsin, treated with L-(tosylamido-2-phenyl) ethyl chloromethyl ketone to inhibit contaminating chymotryptic activity, chemically modified to promote stability and further purified to remove autolysis fragments, resulting in a highly stable trypsin product resistant to autolysis while retaining specificity. Supplied as 1mgP/ml in 1.0mM HCl.
Store at 2-8°C.
Ice Pack required
Code
TRSEQZS
Product details
LS02150
250 ug
$130.00
LS02152
1000 ug
$415.00
LS02155
Bulk
---
Description
Trypsin, Purified, Sequencing Grade II
Source:
Bovine Pancreas
Bovine trypsin that has been treated with L-(tosylamido-2-phenyl) ethyl chloromethyl ketone (TPCK) to inhibit contaminating chymotryptic activity and extensively purified to remove autolysis products. Supplied as a lyophilized powder.
Store at -20°C. PROTECT FROM MOISTURE.
Ice Pack required
Activity
≥150 units per mg protein (at least 8,625 BAEE/2875 USP/NF units per mg protein)
Code
TRSEQII
Product details
LS02115
4x25 µg
$88.00
LS02117
4x100 µg
$235.00
LS02119
1 mg
$420.00
LS02118
Bulk
---
Description
Trypsin, TPCK Treated
Source:
Bovine Pancreas
A chromatographically purified, diafiltered, lyophilized powder that has been treated with L-(tosylamido-2-phenyl) ethyl chloromethyl ketone (TPCK) to inhibit contaminating chymotryptic activity [Kostka, V., and Carpenter, F.: JBC, 239, 1799 (1964)].
Store at 2-8°C. PROTECT FROM MOISTURE.
Activity
≥180 units per mg protein (10,350 BAEE/3,450 USP/NF units per mg protein)
Code
TRTPCK
Product details
LS003740
100 mg
$118.00
LS003741
500 mg
$420.00
LS003744
1 gm
$705.00
LS003742
Bulk
---
Description
Trypsin
Source:
Bovine Pancreas
Supplied as a chromatographically purified, diafiltered and lyophilized powder.
Store at 2-8°C. PROTECT FROM MOISTURE.
Code
TRL3
Product details
LS003708
100 mg
$55.00
LS003707
1 gm
$340.00
LS003709
Bulk
---
Description
Trypsin
Source:
Bovine Pancreas
Supplied as a dialyzed and lyophilized powder.
Store at 2-8°C. PROTECT FROM MOISTURE.
Code
TRL
Product details
LS003702
100 mg
$30.00
LS003703
1 gm
$160.00
LS003704
10 gm
$1,150.00
LS003706
Bulk
---
Description
Trypsin, TPCK-Treated, Irradiated
Source:
Bovine Pancreas
Chromatographically purified trypsin treated with L-(tosylamido-2-phenyl) ethyl chloromethyl ketone (TPCK) to inhibit contaminating chymotryptic activity according to Kostka, V., and Carpenter, F.H.: JBC, 239, 1799 (1964), Code: TRTPCK, lyophilized, irradiated and tested for the absence of mycoplasma and extraneous virus according to 9 CFR 113.53c. Each vial is filled to contain 100 mg.
Store at 2-8°C.
Code
TRTVMF
Product details
LS003750
100 mg
$180.00
LS003752
5x100 mg
$725.00
Description
Trypsin, 2X, Sterile, Irradiated
Source:
Bovine Pancreas
Purified trypsin (Code: TRL), lyophilized, irradiated and tested for the absence of mycoplasma and extraneous virus according to 9 CFR113.53c. Each vial is filled to contain 100 mg.
Store at 2-8°C.
Activity
≥180 units per mg protein (10,350 BAEE/3,450 USP/NF units per mg protein)
Code
TRLVMF
Product details
LS004454
100 mg
$116.00
LS004452
5x100 mg
$420.00
LS004458
Bulk
---
Description
Trypsin, 0.22µ Filtered
Source:
Bovine Pancreas
Trypsin chromatographically purified, diafiltered (Code TRL3) filtered thorugh a 0.22 micron pore size membrane and lyophilized in sterile vials. This product is not tested for pyrogenicity.
Store at 2-8°C.
Activity
≥180 units per mg protein (10,350 BAEE/3,450 USP/NF units per mg protein)
Code
TRLS
Product details
LS003736
50 mg
$54.00
LS003734
5x50 mg
$235.00
LS003738
Bulk
---