Quick Human GPC3 (Glypican 3) ELISA Kit

Catalog Number:

V90R75

Background:

Human Glypican-3 (GPC3) is a membrane-bound protein highly expressed in hepatocellular carcinoma (HCC) but absent in normal adult liver tissue, making it a key biomarker and therapeutic target.

Applications:

This kit is developed to quickly titrate human GPC3 sample in high throughput. It is for research use only.

Description:

These kits were validated using human GPC3 protein (see figure). The detection linear range is 0.01 – 50 µg/ml; Sensitivity is 0.01 µg/ml.

Biosafety Level:

BSL-1

Organism:

Human Glypican-3 (GPC3)

Assay Principles (Take neutralizing antibody as example):

This kit is developed for a sandwich ELISA format to specifically capture human GPC3 protein in serum, plasma, tissue homogenates, cell culture supernatant and other biological fluids. The whole process should take about 2.5 hours. A molecule specific for GPC3 has been pre-coated onto a detachable 96-well microtiter plate. The standards and test samples are added into the wells and the GPC3 present in each sample is bound to the wells by the immobilized molecule. Following incubation, the wells are washed and then incubated with HRP labeled detector antibody, which binds the captured GPC3 present in each well. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-colored product that changes to yellow after addition of stop solution. The density of yellow is proportional to the amount of GPC3 captured in each well. The concentration of GPC3 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.

The kit consists of detachable 96-well pre-coated plate, GPC3 protein standard, HRP labeled detector antibody, TMB and sample dilution/wash/stop buffer.

Brief protocol (detailed protocol is provided with the product):

1. Add your samples containing human GPC3 and provided purified GPC3 protein standard into pre-coated plate (or strips based on sample size). Incubate at 37°C for 1 hour.
2. Wash wells. Add HRP labeled detector antibody to wells. Incubate at room temperature for 1 hour.
3. Wash wells. Add TMB and stop solution. Read absorbance (OD) at 450 nm. The OD Value reflects the amount of GPC3 in the samples, which is then quantified against GPC3 standard curve to calculate titer.

Storage and shipping condition:

Shipment with room temperature. The unopened kit is stable for 6 months from the date of manufacture when stored at 2°C to 8°C. Avoid light. The shelf life is 60 days from the date of opening.