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Lactate Dehydrogenase (LDH) Activity Assay Kit (WST-8 method)– MSE Supplies LLC

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Lactate Dehydrogenase (LDH) Activity Assay Kit (WST-8 method)

SKU: E-BC-K766-M-96

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Lactate Dehydrogenase (LDH) Activity Assay Kit (WST-8 method)

SKU # E-BC-K766-M
Detection Instrument Microplate reader (450 nm)
Detection method: Colorimetric method


Product Details

Properties

Synonyms LDH
Sample type Serum, plasma, animal tissue, hydrothorax, cells, cell culture supernatant
Sensitivity 0.11 U/L
Detection range 0.11-39.9 U/L
Detection method Colorimetric method
Assay type Enzyme Activity
Assay time 30 min
Precision Average inter-assay CV: 2.300% | Average intra-assay CV: 2.300%
Other instruments required Microplate, Micropipettor, Vortex mixer, Water bath
Storage -20℃
Valid period 12 months


Images

Q Li et al investigate the hypoxia-induced neurogenic bladder fibrosis. Lactate dehydrogenase (LDH) activity of mouse and human serum was determined using LDH activity assay kit (E-BC-K766-M).

LDH activity in serum was significantly increased by NB. (**P<0.01, ***P<0.001)

 

Dilution of Sample

 

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.11-39.9 U/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
Human serum 10-20
Dog serum 10-20
Mouse serum 50-100
Cynomolgus monkey serum 10-20
10% Rat spleen tissue homogenate 150-250
10% Rat liver tissue homogenate 250-350
10% Rat kidney tissue homogenate 250-350
10% Rat lung tissue homogenate 250-350

 

Note: The diluent is reagent 1.

 

Detection Principle

Lactate dehydrogenase (LDH) is an oxidoreductase. LDH catalyzes the conversion of lactate to pyruvic acid and back, as it converts NAD+ to NADH and back. LDH is composed of four subunits (tetramer). The two most common subunits are the LDH-M and LDH-H protein. LDH is released into the blood by cells after tissue damage or erythrocyte hemolysis. Extracellular LDH activity is used to detect cell damage or cell death.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Lysis Solution 60 mL × 1 vial 60 mL × 2 vials -20℃, 12 months
Reagent 2 Substrate 1.5 mL ×1 vial 1.5 mL × 2 vials -20℃, 12 months
Reagent 3 Chromogenic Agent 1.5 mL ×1 vial 1.5 mL × 2 vials -20℃, 12 months,
shading light
Reagent 4 Coenzyme Powder × 1 vial Powder × 1 vial -20℃, 12 months
Reagent 5 Stop Solution 3 mL ×1 vial 6 mL ×1 vial -20℃, 12 months
Reagent 6 NADH Standard Powder × 1 vial Powder × 1 vial -20℃, 12 months
Microplate 96 wells No requirement
Plate Sealer 2 pieces

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).

Parameters Sample 1 Sample 2 Sample 3
Mean (U/L) 2.60 18.50 26.00
%CV 2.6 2.3 2.0


Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (U/L) 2.60 18.50 26.00
%CV 2.4 2.5 2.0


Sensitivity

The analytical sensitivity of the assay is 0.11 U/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.

 

Standard Curve

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:

Concentration (μmol/L) 0 50 100 150 200 250 300 400
Average OD 0.054 0.171 0.292 0.430 0.559 0.691 0.845 1.114
Absoluted OD 0.000 0.117 0.238 0.377 0.505 0.638 0.792 1.061