HA-tag Rapid Assay (Colloidal Gold), 10 T

 Product Information

Product Description/Introduction

It is a semi-quantitative HA protein assay card based on colloidal gold lateral chromatography and the principle of competitive assay for rapid detection of HA tagged proteins in cell culture supernatants, cell/bacterial lysate or after protein purification.

The product mainly consists of a sample pad, binding pad, chromatography membrane, absorbent paper, and backing plate.Goat anti-mouse IgG polyclonal antibody (Control Line) and HA-tagged protein (Test Line) were immobilized on the chromatographic membrane; colloidal gold-labeled HA-tagged antibody was immobilized on the binding pad. When a drop of the sample to be measured is added to the sample pad, the liquid moves toward the binding pad by capillary action. If the sample does not contain HA-tagged protein, the gold-labeled anti-HA-tagged antibody on the conjugate pad binds to the HA-tagged protein immobilized at the Test Line to form an "Au-anti-HA-tagged antibody-HA-tagged protein" complex, resulting in a red line at the Test Line; if the tested sample contains HA-tagged protein, the HA-tagged protein binds to the gold standard anti-HA-tagged antibody on the binding pad to form an "Au-anti-HA-tagged antibody-HA-tagged protein" complex, and the unreacted gold standard anti-HA-tagged antibody on the binding pad reacts with the HA-tagged protein on the Test Line, a red line appears on the Test Line as the content of HA-tagged protein increases, the coloration of the Test Line gradually decreases until it is completely colorless. Regardless of the presence of HA-tagged proteins in the sample being examined, the gold-labeled anti-HA-tagged antibody on the binding pad will be chromatographed upward to the control line, where it reacts with the goat anti-mouse IgG to appear as a red line. The red line at control line is the criterion for determining whether the chromatography process is normal.

Storage and Shipping Conditions

Ship and store at room temperature, valid for 12 months.

Product Components

Assay Procedures/Protocols

1. Open the package to take out the detection card and lay it flat on the lab bench.

2. Pipette 80 µL of culture supernatant, cell/bacterial lysis product, or post-protein purification sample dropwise into the sample hole.

3. Wait 8-10 minutes for the color bands to appear, it is invalid after 15 minutes.

Results Judgement

1. Negative result: Both the control line and the test line are coloured, with the test line showing the darkest colour.

2. Positive result: The control line is coloured, the test line is weakly coloured or not coloured.

3. Invalid result: When the control line is not coloured, the test should be repeated whether the test line is coloured.

Note: When the sample does not contain HA-tag protein, the color of the test line is darkest, and gradually decreases as the amount of HA-tag protein in the sample increases, until the colour disappears.

Results Display

Take out the detection card and lay it flat on the lab bench. Dilute the Flag-tagged protein with PBS to 10 μg/mL, 20 μg/mL and 50 μg/mL respectively, and then add 100 μL of dilution to the sample hole and leave it for 8 min, observe the results.

Results of HA-tag Rapid Assay(colloidal gold) for 8 min

Sensitivity

The lower limit of detection is 20 μg/mL, when the HA-tag protein content in a sample is below 20 μg/mL, it is not easily distinguish with the naked eye.

Note

1. If the colour of the sample causes a high background, the sample can be diluted with PBS or culture medium before chromatography.

2. For the accuracy of the results, a negative control and positive control should be set up, and it is recommended that the control sample is the same matrix as the sample to be tested.

3. This test card is not recommended for freezing.

4. Once opened please use quickly.

For Research Use Only!

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