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
Sample type | Limit Of Quantitation (ng/g ) |
Fresh milk | 0.03 |
Milk powder | 0.1 |
Yogurt | 0.05 |
Whey powder | 0.1 |
Condensed milk | 0.15 |
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:
Incubate at 50°C -55°C for 3 hours. Vortex the sample for 5 seconds
every hour during the incubation.
