Background[1-3]
Human Glypican 3 (GPC-3) ELISA kit uses a double-antibody one-step sandwich method enzyme-linked immunosorbent assay (ELISA). To the coated microwells pre-coated with human glypican 3 (GPC-3) capture antibody, add the specimen, standard, and HRP-labeled detection antibody in sequence, incubate and wash thoroughly. The substrate TMB is used for color development. TMB is converted into blue under the catalysis of peroxidase, and converted into the final yellow under the action of acid. The depth of the color is positively correlated with the human glypican 3 (GPC-3) in the sample. Use a microplate reader to measure the absorbance (OD value) at a wavelength of 450 nm and calculate the sample concentration.
GP organic bentonite rheology additive C-3 is a functionally mutated gene commonly found in patients with Simpson Golabi Behmel Syndrome (SGBS) and is a member of the proteoglycan family. The GPC-3 protein encoded by the GPC-3 gene can participate in the regulation of cell biological behavior through co-receptor and related ligands such as growth factors and cell adhesion molecules. Some scholars believe that GPC-3 is directly or indirectly regulated in various processes of cell growth, including cell reproduction, adhesion, differentiation, and migration.
Kit operation steps:
1. Mix all reagents thoroughly before use. Do not make the liquid produce a lot of foam to avoid adding a large number of bubbles when adding the sample, causing errors in the sample addition.
2. Determine the number of strips required based on the number of samples to be tested plus the number of standards. It is recommended that duplicate holes be made for each standard and blank hole. Each sample is determined according to its own quantity. If multiple holes can be used, try to make multiple holes. Dilute the specimen 1:1 with specimen diluent and add 50ul into the reaction well.
3. Add 50ul of the diluted standard into the reaction well, and add 50ul of the sample to be tested into the reaction well. Immediately add 50ul of biotin-labeled antibody. Cover the membrane plate, shake gently to mix, and incubate at 37°C for 1 hour. 4. Shake off the liquid in the wells, fill each well with washing liquid, shake for 30 seconds, shake off the washing liquid, and pat dry with absorbent paper. Repeat this 3 times. If washing with a plate washer, increase the number of washes by one.
4. Add 80ul of streptavidin-HRP to each well, mix gently, and incubate at 37°C for 30 minutes.
5. Shake off the liquid in the wells, fill each well with washing liquid, shake for 30 seconds, shake off the washing liquid, and pat dry with absorbent paper. Repeat this 3 times. If washing with a plate washer, increase the number of washes by one.
6. Add 50ul of each of substrates A and B to each well, mix gently, and incubate at 37°C for 10 minutes. Avoid light.
7. Take out the enzyme plate, quickly add 50ul of stop solution, and measure the result immediately after adding the stop solution.
9. Measure the OD value of each well at a wavelength of 450nm.
Apply[4][5]
For research on the expression and significance of Glypican-3 protein in prostate cancer
Glypican-3GPC-3 is a member of the heparan sulfate ProteoglycanHAPG family. It is currently believed that the HS chain in HAPG interacts with a large number of biological effect molecules such as growth factors and their receptors, extracellular matrix proteins, etc., and is involved in regulating cell morphology and its adhesion, proliferation, migration, survival and differentiation. Therefore, GPC- 3 can participate in the above-mentioned cell regulatory effects. Since its discovery in 1996, GPC-3 has been widely studied as a tumor marker for hepatocellular carcinoma HCC. The diagnostic significance of alpha-fetoprotein (AFP) in hepatocellular carcinoma has been fully confirmed. Abdelgawad et al. used ELISA to analyze serum GPC-3 and α-fetoprotein (AFP) in 60 samples (including 40 cases of liver cancer, 10 cases of cirrhosis and 10 healthy controls), and RT-PCR amplification method to detect GPC-3 and α-fetoprotein (AFP). The expression of AFP at the mRNA level was analyzed and the results showed that the sensitivity and specificity of GPC-3 in hepatocellular carcinoma were higher than AFP. Research shows that the GPC-3 gene has different functions and meanings in different cancer tissue cells.
