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Papain - Assay
Source:
Carica papaya Latex
CAS:
9001-73-4
EC:
3.4.22.2
Papain Assay
Method
A titrimetric determinatin of the acid produced during the hydrolysis of benzoyl-L-arginine ethyl ester (BAEE). One unit will hydrolyze one micromole of benzoyl-L-arginine ethyl ester per minute at 25°C and pH 6.2 under the specified conditions.
Reagents
Enzyme diluent (Activation buffer): Prepare fresh daily by mixing the following:
0.01 M EDTA 10 ml 0.06 M Mercaptoethanol 0.1 ml 0.05 M Cysteine⋅HCl 10 ml Reagent grade water 70 ml Substrate solution: Prepare fresh daily by mixing the following:
0.058 M BAEE 15.0 ml 0.01 M EDTA 0.8 ml 0.05 M Cysteine⋅HCl 0.8 ml - Adjust pH to 6.2 and dilute to a final volume of 21 ml with reagent grade water.
- Titrant: 0.01-0.02 N NaOH, standardized
Enzyme
Activate enzyme by dissolving in enzyme diluent to a concentration of 0.05-0.1 mg/ml. Under these conditions activation is complete within 30 minutes.
Determination of protein concentration
Procedure
The reaction can be measured with either an automatic titrator or a laboratory pH meter. The titration vessel should be maintained at 25°C.
Pipette the following into the titration vessel at 25°C:
Substrate solution 5.0 ml 3.0 M NaCl 5.0 ml Reagent grade water 5.0 ml
At zero time add 0.1 ml of appropriately diluted enzyme and adjust the pH to 6.2. Record the amount of standardized NaOH added per minute to maintain the pH at 6.2 after a constant rate is achieved.
Calculation
Papain Products
Description
Activity
Code
Cat. #
Size
Price
Description
Papain, Suspension
Source:
Carica papaya Latex
Supplied as a two times crystalline suspension in 50mM sodium acetate, pH 4.5. To ensure full activity, the enzyme should be incubated in a solution containing 1.1mM EDTA, 0.067mM mercaptoethanol and 5.5mM cysteine-HCl for 30 minutes. It is recommended that the enzyme be 0.22 micron filtered after dissolution and prior to use.
Store at 2-8°C.
Ice Pack required
Code
PAP
Product details
LS003124
25 mg
$44.00
LS003126
100 mg
$94.00
LS003127
1 gm
$625.00
LS003128
Bulk
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Description
Papain, Lyophilized
Source:
Carica papaya Latex
Supplied as a lyophilized powder prepared from a two times crystalline suspension, Code: PAP. To ensure full activity, the enzyme should be incubated in a solution containing 1.1mM EDTA, 0.067mM mercaptoethanol and 5.5mM cysteine-HCl for 30 minutes. It is recommended that the enzyme be 0.22 micron filtered after dissolution and prior to use.
Store at 2-8°C.
Code
PAPL
Product details
LS003118
25 mg
$47.00
LS003119
100 mg
$105.00
LS003120
1 gm
$720.00
LS003122
Bulk
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Description
PDS Kit, Papain Vial
Source:
Papaya Latex
A component of the Papain Dissociation System, for use in the tissue dissociation method of Huettner, J.E., and Baughman, R.W.: J. Neuroscience, 6, 3044 (1986). Contains papain, L-cysteine, and EDTA. This material is 0.22 micron membrane filtered and lyophilized in autoclaved vials. A vial reconstituted with 5 ml of EBSS or equivalent yields a solution at 20 units of papain per ml in 1mM L-cysteine with 0.5mM EDTA.
Store at 2-8°C.
Code
PAP2
Product details
LK003176
1 vi
$31.00
LK003178
5 vi
$104.00