Adenosine Deaminase Assay

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Adenosine Deaminase Assay

Method

The reaction velocity is measured by a change in absorbance at 265 nm resulting from the deamination of adenosine. One unit converts one micromole of adenosine to inosine per minute at 25°C and pH 7.4 under the specified conditions.

Reagents

1.4 mM Adenosine in 0.05 M Phosphate buffer, pH 7.4
0.05 M Phosphate buffer, pH 7.4

Enzyme

Dilute immediately before use in ice cold 0.05 M phosphate buffer, pH 7.4 to a concentration of 0.2 - 0.8 units / ml.

Procedure

Adjust spectrophotometer to 265 nm and 25°C.

Pipette into cuvettes as follows:

0.05 M Phosphate buffer, pH 7.4 2.88 ml

1.4 mM Adenosine 0.1 ml

Incubate in spectrophotometer for 3-5 minutes to achieve temperature equilibration and establish blank rate, if any. At zero time add 0.02 ml enzyme solution and mix thoroughly. Record decrease in A₂₆₅ for 2-3 minutes. Use the initial rate to calculate ΔA₂₆₅/min.

Calculation

\frac{\text{Units}}{\text{mg}} = \frac{\frac{\Delta \text{A}_{265}}{\text{min}}}{\frac{\text{8.1 x mg enzyme}}{\text{ml reaction mixture}}}

Adenosine Deaminase Products

Description

Adenosine Deaminase

Source:

Calf Spleen

A chromatographically purified, dialyzed, lyophilized powder. Prepared by a modification of the method of Pfrogner, Arch. Biochim. Biophys., 119, 141 (1967).

Store at 2-8°C.

…

Activity

≥15 units per mg dry weight

Certificate of Analysis

Code

ADA

Product details

LS009043

250 un

$210.00

LS009044

Bulk

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0.05 M Phosphate buffer, pH 7.4	2.88 ml
1.4 mM Adenosine	0.1 ml

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Adenosine Deaminase - Assay

Adenosine Deaminase Assay

Adenosine Deaminase Products