Colloidal gold test paper strip for semiquantitatively detecting concentration of GFAP (Glial Fibrillary Acidic Protein) in human serum
A semi-quantitative detection and colloidal gold test paper technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome detection methods, shortened detection time, and long detection time, and achieve high specificity, low cost, and high sensitivity Effect
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Embodiment 1
[0030] Step 1: Preparation of detection membrane
[0031] Dilute the anti-GFAP capture monoclonal antibody to four different concentrations with PBS buffer, and spot on the nitrocellulose membrane with a film spotter from high to low, each line is 20 μl, and the line width is 1-1.5 mm. The line spacing is 5mm. Its coating amount means that 2ng / ml, 4ng / ml, 8ng / ml, and 16ng / ml can be detected. In the same way, spot 200 μg / ml goat anti-mouse IgG after the test line as a quality control line with a film spotter, with a line width of 1-1.5 mm and a distance of 8 mm from the nearest test line. Block with PBS buffer containing 1% BSA for 2 hours, dry in the shade at room temperature, and store at 4°C.
[0032] Step 2 Preparation of Colloidal Gold Bonded Film
[0033] 1) Anti-GFAP antibody
[0034] Monoclonal antibodies were purchased from Santa Cruz Biotechnology.
[0035] 2) Preparation of anti-GFAP antibody-colloidal gold complex
[0036] Colloidal gold with a diameter of 20nm ...
Embodiment 2
[0043] Application of colloidal gold test strips for semi-quantitative detection of GFAP concentration in human serum
[0044] (1) Detect GFAP concentration in human serum with test strips
[0045] Use a pipette to draw 120 μl of serum sample, and drop it vertically onto the sample loading site of the colloidal gold test strip. Start timing, wait for the red band to appear, and compare it with the standard color card within 10 minutes. If it exceeds 20 minutes, the result will be invalid.
[0046] (2) Interpretation of results
[0047] The four concentration gradient marking lines (ie detection lines) involved from top to bottom in this colloidal gold test strip are: 16ng / ml, 8ng / ml, 4ng / ml, 2ng / ml. The test results are explained as follows:
[0048] When only one quality control line appears, it means that the GFAP concentration is less than 2ng / ml.
[0049] When two lines containing quality control lines appear, it means that the concentration of GFAP is between 2ng / ml a...
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