5'-nucleotidase diagnosis reagent kit and 5'-nucleotidase active concentration determination method
A technology of nucleotidase and nucleoside monophosphate, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, color/spectral characteristic measurement, etc., and can solve problems such as difficulty in popularization and application, strong acid pollution of the environment, and complicated methods
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Embodiment 1
[0042] The 5'-nucleotidase diagnostic reagent of this embodiment is a single reagent, comprising:
[0043] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L
[0044] Stabilizer 500mmol / L
[0045] Reduced coenzyme 0.25mmol / L
[0046] Pyruvate carboxylase 6000U / L
[0047] Formaldehyde dehydrogenase 10000U / L
[0048] Nucleoside monophosphate 8mmol / L
[0049] Adenosine diphosphate 6mmol / L
[0050] Oxaloacetate 12mmol / L
[0051] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.
[0052] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, the volume of the tested 5'-nucleotidase sample and reagent The ratio is 1 / 25, the reaction direction is negative reaction (decreasing reaction), the delay time is about ...
Embodiment 2
[0055] The 5'-nucleotidase diagnostic reagent of this embodiment is a double reagent, comprising:
[0056] Reagent 1
[0057] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L
[0058] Stabilizer 50mmol / L
[0059] Reduced coenzyme 0.25mmol / L
[0060] Nucleoside monophosphate 8mmol / L
[0061] Adenosine diphosphate 6mmol / L
[0062] Oxaloacetate 12mmol / L
[0063] Reagent 2
[0064] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L
[0065] Stabilizer 500mmol / L
[0066] Pyruvate carboxylase 6000U / L
[0067] Formaldehyde dehydrogenase 10000U / L
[0068] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.
[0069] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, tested 5'-nucleotidase sample and reagent 1, The volume ratio of rea...
Embodiment 3
[0072] The 5'-nucleotidase diagnostic reagent of the present embodiment is three reagents, comprising:
[0073] Reagent 1
[0074] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L
[0075] Stabilizer 50mmol / L
[0076] Reduced coenzyme 0.25mmol / L
[0077] Nucleoside monophosphate 8mmol / L
[0078] Adenosine diphosphate 6mmol / L
[0079] Oxaloacetate 12mmol / L
[0080] Reagent 2
[0081] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L
[0082] Stabilizer 500mmol / L
[0083] Formaldehyde dehydrogenase 10000U / L
[0084] Reagent 3
[0085] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L
[0086] Stabilizer 500mmol / L
[0087] Pyruvate carboxylase 6000U / L
[0088] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.
[0089] When measuring the concentration of 5'-nucleotidase activity, set it on the automatic biochemical analyzer: tem...
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