High Reproducibility Drug Residue Test Kit Chloramphenicol (CAP) ELISA Test Kit

Chloramphenicol (CAP) ELISA Test Kit

Product Description

REAGEN™Chloramphenicol (CAP) ELISA Test Kit is a competitive enzyme immunoassay for the quantitative analysis of chloramphenicol in fish, shrimp, eggs, honey, meat (beef, chicken and pork), milk, milk powder , condensed milk and serum.

The unique features of the kit are:

1.High recovery (80-105%), rapid (10 - 40 minutes), and cost-effective extraction methods.

2.Detection limit is 0.05 ng/g .

3.High reproducibility.

4.A quick ELISA assay (less than1 hour regardless of number of samples).

Procedure Overview

The method is based on a competitive colorimetric ELISA assay. The drug of interest has been coated in the plate wells. During the analysis, sample and the CAP antibody are added along with secondary antibody, tagged with a peroxidase enzyme. If the CAP residue is present in the sample, it will compete for the CAP antibody, thereby preventing the antibody from binding to the CAP attached to the well. The resulting color intensity, after addition of the HRP substrate (TMB), has an inverse relationship with the CAP residue concentration in the sample.

Kit Contents, Storage and Shelf Life

REAGEN™Chloramphenicol (CAP) ELISA Test Kit has the capacity for 96 determinations or testing of 42 samples in duplicate (assuming 12 wells for standards). Return any unused microwells to the foil bag and reseal them with the desiccant provided in the original package.Store the kit at 2-8°C *. The shelf life is 12 months when the kit is properly stored.      

Detection Limit Of Quantitation

Be sure samples are properly stored. In general, samples should be refrigerated at 2-4°C for no more than 1-2 days. Freeze samples to a minimum of -20°C if they need to be stored for a longer period. Frozen samples can be thawed at room temps (20 – 25°C / 68 – 77°F) or in a refrigerator before use.

Preparation of 1X Sample Extraction Buffer:

Mix 1 volume of 10X Sample Extraction Buffer with 9 volumes of distilled water.

Fish/Shrimp/Meat (Beef/Chicken/Pork)/Egg.

A:

• Homogenize a reasonable amount of sample with a suitable mixer.

• Weight out 3g of the homogenized sample and mix with 6 mL of ethyl acetate, vortex 3 min at max speed.

• Centrifuge for 5 minutes at 4,000 x g at room temperature (20 – 25°C / 68 – 77°F).

• Transfer 4 mL of the ethyl acetate supernatant (corresponding to 2g of the original sample) into a new vial and use a rotary evaporator to dry the sample in a 60-70°C water bath under reduced pressure. Alternatively, the sample can be dried by blowing nitrogen gas in a 60-70°C water bath.

• Dissolve the dried residue in 2 mL of n-hexane.

• Add 1 mL of 1X Sample Extraction Buffer and mix by vortexing at maximum speed for 2 minutes.

• Centrifuge for 10 minutes at 4,000 x g at room temp. Discard the upper hexane layer.

• Use 50 mL of the lower aqueous layer per well for the assay. In case emulsion happened, remove the upper n-hexane layer and incubate the lower aqueous layer in water bath for 3 min at 80-95°C.

Note: Dilution factor: 0.5.

Fish/Shrimp/Meat (Beef/Chicken/Pork)/Egg.

B:

• Mix 1 g of the homogenized sample with 0.5 mL of 1X Sample Extraction Buffer, 3.5 mL of distilled water, 0.5 mL of 1 M HCl and 20 mL of 50 mM 2-Nitrobenzaldehyde by vortexing for 30 seconds.

Incubate at 50°C -55°C for 3 hours. Vortex the sample for 5 seconds every hour during the incubation.