This test kit is based on the competitive enzyme immunoassay for the detection of Gentamicin
in the sample. The coupling antigen is pre-coated on the micro-well stripes. The Gentamicin in the sample and pre-coated coupling antigen on the micro-well stripes compete for the anti-Gentamicin antibody. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the sample has a negative correlation with the Gentamicin in it. The value is compared to the standard curve and the Gentamicin concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 0.1 ppb
Chicken 4 ppb
Liver 4 ppb
1 Bring all reagents and micro-well strips to the room temperature (20-25 ℃) before use.
2 Return all reagents to 2-8 ℃ immediately after use.
3 The reproducibility of the ELISA analysis, to a large degree, depends on the consistency of plate washing. The correct operation of plate washing is the key point in the procedures of ELISA.
4 For the incubation at constant temperatures, all the samples and reagents must avoid light exposure, and each microplate should be sealed by the cover membrane.
6.2 Operation procedures
1. Take out the kit from the refrigerated environment. Take out all the necessary reagents from the kit and place at the room temperature (20-25 ℃) for at least 30 min. Note that each liquid reagent must be shaken to mix evenly before use.
2. Take the required micro-well strips and plate frames. Re-sealed the unused microplate, stored at 2-8 ℃, not frozen.
3. Solution preparation: dilute 40 mL of the concentrated washing buffer (20× concentrated) with the distilled or deionized water to 800 mL (or just to the required volume) for use.
4. Numbering: number the micro-wells according to samples and standard solution; each sample and standard solution should be performed in duplicate; record their positions.
5. Add 50 µL of the sample or standard solution to separate duplicate wells, and add 50 µL enzyme conjugate and then 50 µL of the antibody solution into each well. Vortex evenly, seal the microplate with the cover membrane, and incubate at 25 ℃ for 1 h
6. Pour liqiud out of the microwells, add 250 µL/well of washing buffer for 10 sec, repeat four to five times, then flap to dry with absorbent paper (if there are the bubbles after flapping, cut them with the clean tips).
7. Coloration: add 50 µL of the substrate A solution and then 50 µL of the B solution into each well. Mix gently by shaking the plate manually, and incubate at 25 ℃ for 15 min at dark for coloration;
8. Determination: add 50 µL of the stop solution into each well. Mix gently by shaking the plate manually. Set the wavelength of the microplate reader at 450 nm to determine the OD value (Recommend to read the OD value at the dual-wavelength 450/630 nm within 5 min).
Contact Name: Jiang Yongqing
Company Name: Shenzhen Lvshiyuan Biotechnology Co, Ltd
Street Address: Rm.507, No.2,
Longgang Overseas Venture Park,
Member name: Shenzhenlvshiyuanbiotec
Member Since: 07 March 2008
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