Rapid Detection Kit for V5-tagged Proteins (Competitive Assay)

The Rapid Detection Kit is used for quick, simple detection of V5-tagged proteins through a colloidal gold-based lateral flow immunoassay (LFIA). The LFIA strip is composed of membrane immobilized antibodies, sample pad, conjugate pad, and absorbent pad.
The gold conjugated monoclonal antibodies against V5-tag are immobilized at conjugate pad. The V5-tagged proteins are immobilized over test line. The secondary antibodies are immobilized at control zone. The sample is applied to the sample pad at one end of the strip and it subsequently migrates to the other parts of strip by capillary action. If the sample contains proteins with V5-tag, the gold conjugated capture antibodies will bind to these V5-tagged proteins in the sample and not to the immobilized V5-tagged proteins in the test zone. Intensity of color at test line inversely corresponds to the concentration of V5-tagged proteins in the sample. In a sample where no V5-tagged proteins are present or the concentration of V5-tagged proteins is below detectable levels, the gold-conjugated capture antibodies will bind to the V5-tagged proteins embedded on the test line and form a red line, representing a negative result. A red control line will appear in both positive and negative results. In both positive and negative tests, the gold-conjugated anti-V5 antibodies will bind to the secondary antibodies immobilized at the control line.
The recommended working range of the competitive lateral flow assay is 2 ug/ml to 18 ug/ml of purified V5-tagged proteins. The color of the test line is negatively related with the concentration of V5-tagged proteins in the sample. When the concentration of V5-tagged proteins in the sample reaches 2 ug/ml and above, the intensity of the test line will be effectively weakened, showing a light red band or no color. The lateral flow strip will continue to show positive results at concentrations greater than 18 ug/ml, visualized by complete loss of the test line.

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