Intended Use
The Reagen Malachite green plate kit is a competitive elisa for the quantitative analysis of Malachite green in aquatic products.
Assay Principles
The Reagen Malachite green plate kit is a competitive enzyme-labeled immunoassay. Malachite green is extracted from a sample by blending or shaking with extraction solvent. The diluted Malachite green sample extract and calibrators are pipetted into the test wells followed by malachite green antibody into the test wells to initiate the reaction. After this 30 minute incubation the wells are washed with laboratory grade water. Add Malachite green HRP conjugate for 30 minutes. Following a second wash after the incubation, a clear substrate is added to the wells and any bound enzyme conjugate causes the conversion to a blue color. After the 15 minute incubation, the reaction is stopped and the amount of color in each well is read . The color of the unknown samples is compared is compared to the color of the calibrators and the Malachite green concentration of the samples is derived.
Detection Linmit
Aquatic products: 0.5ppb
Reagents And Materials Provided
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Kit Contents
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Amount
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Storage
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Malachite green Coated Plate
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1 x 96-well Plate
(8 wells x 12 strips)
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2-8°C
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Malachite green Standards:
Negative control (white cap tube)
0.015 ng/mL (yellow cap tube)
0.05 ng/mL (orange cap tube)
0.15 ng/mL (pink cap tube)
0.5ng/mL (purple cap tube)
1.5ng/mL (blue cap tube)
50 ng/mL (spiking, optional, red cap tube)
|
0.8 mL
0.8 mL
0.8 mL
0.8 mL
0.8 mL
0.8 mL
0.4 mL
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2-8°C *
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Malachite green Antibody
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8 mL
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2-8°C *
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100X HRP Conjugate
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100 µL
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2-8°C *
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HRP Conjugate Diluent
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20 mL
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2-8°C
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20X Wash Solution
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28 mL
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2-8°C
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Stop Buffer (Caution 1N HCI . Handle with care.)
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12mL
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2-8°C
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TMB Substrate
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12 mL
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2-8°C
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10 X Sample Dilution Buffer
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25 mL
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2-8°C
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Reagent A(for Sample preparation)
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20 g
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2-8°C
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10X Reagent B(used for oxidation LMG to MG) )
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1.8 mL
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2-8°C
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Specificity
The Reagen Malachite green Plate Kit can not differentiate between the various Malachite greens metabolites, but detects their presence to differing degrees. The following table shows the % cross reactivity of Leucomalachite versus Malachite green.