Rat

0.25ng/mL - 8ng/mL

0.1ng/mL

It is used to detect the concentration of rat anti-glomerular basement membrane antibodies (GBM-Ab) in samples such as serum, plasma, cell culture supernatant and tissue.

This kit uses a double antibody sandwich enzyme-linked immunosorbent assay (ELISA). In a microwell ELISA plate pre-coated with anti-rat GBM-Ab antibody (solid phase antibody), rat GBM-Ab calibrators and samples to be tested are added, and then HRP-labeled anti-rat GBM-Ab antibody (enzyme-labeled antibody) is added. After incubation and sufficient washing, unbound components are removed, and a sandwich complex of solid phase antibody-antigen-enzyme-labeled antibody is formed on the solid surface of the microwell plate. Substrates A and B are added, and the substrates produce blue products under the catalysis of HRP. Under the action of the stop solution (acidic solution), they are finally converted into yellow. The absorbance (OD value) is measured at a wavelength of 450nm on an ELISA instrument. The absorbance (OD value) is positively correlated with the concentration of rat GBM-Ab in the sample to be tested. The concentration of rat GBM-Ab in the sample can be calculated by fitting the calibrator curve.

This kit recognizes natural rat anti-glomerular basement membrane antibody (GBM-Ab) and has no cross-reactivity with structural analogs.

See the instruction manual for details

The intra-batch coefficient of variation CV% was less than 10%; the inter-batch coefficient of variation CV% was less than 15%.

The recovery rate is between 85%-115%.

1. ELISA reader capable of detecting absorbance at 450 nm 2. Pipette and tip, sample loading tank 3. 37°C thermostat or water bath 4. Test tubes, centrifuge tubes, measuring cylinders, etc. for preparing reagents 5. Distilled water or deionized water

All reagents and components should be restored to room temperature. For standards, quality control products and samples, duplicate wells are recommended. 1. Prepare the working solutions of various components of the kit according to the method described in the previous instructions. 2. Take out the required strips from the aluminum foil bag, seal the remaining strips with self-sealing bags and put them back in the refrigerator. 3. Set up standard wells and sample wells, and add 50μL of different concentrations of standards to each standard well; 4. Add 50μL of the sample to be tested to the sample well; do not add to the blank well. 5. Except for the blank well, add 100μL of horseradish peroxidase (HRP) labeled detection antibody to each well of the standard well and sample well, seal the reaction well with a sealing film, and incubate at 37℃ water bath or constant temperature box for 60min. 6. Discard the liquid, pat dry on absorbent paper, fill each well with washing solution (350μL), let it stand for 1min, shake off the washing solution, pat dry on absorbent paper, and repeat the washing process 5 times (you can also use a plate washer to wash the plate). (Tip: To obtain ideal experimental results, the residual liquid must be completely removed. After washing, please proceed to the next step immediately and do not let the microplate dry.) 7. Add 50μL of substrate A and B to each well and incubate at 37℃ in the dark for 15 minutes. 8. Add 50μL of stop solution to each well and measure the OD value of each well at a wavelength of 450nm within 15 minutes.