Method and appts. for selective biological material detection
A bioassay and biological activity technology, applied in the field of detection of pathogenic microorganisms or biological substances, can solve problems such as unexpected sensors
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Embodiment 1
[0056] Example 1 Antibody Detection on the Surface of Pretreated Thin Layer Polyethylene Foils:
[0057] 0.1M Sodium Carbonate (Na 2 CO 3 )-sodium bicarbonate (NaHCO 3 ) buffer to dilute rabbit polyclonal IgG to a final concentration of 2.0 μg / ml.
[0058] Using a 2"x3" grid, 75 μL (150 ng) was applied to the pretreated polyethylene sheet with a 1" gap.
[0059] The antibody-treated polyethylene sheets were dried at 37°C for 1.5 hours.
[0060] The dried film was washed 3 times with pH 7.4 phosphate buffer.
[0061] In 1% casein, 0.1M potassium ferricyanide K 3 Fe(CN) 6 , 0.1% phosphate glass (Na 15 P 13 o 40 -Na 20 P 18 o 55 ) diluted goat anti-rabbit IgG conjugated to HRP (GαR HRP ) to a concentration of 1:7000, pH7.4.
[0062] Add 125 μL of diluted G with a precision pipette HRP into the grid-backed polyethylene film at 1" intervals, coinciding with where previously covered by RαG.
[0063] The flakes were incubated for 30 minutes at room temperature.
[00...
Embodiment 2
[0068] Example 2 Whole Sandwich Immunoassay on the Surface of Pretreated Thin Layer Polyethylene Foils
[0069] 0.1M Sodium Carbonate (Na 2 CO 3 )-sodium bicarbonate (NaHCO 3 ) buffer to dilute rabbit polyclonal IgG to a final concentration of 2.0 μg / ml.
[0070] A 13 x 9 cm pretreated thin layer polyethylene sheet from DuPont was inserted into a BIO-RAD DOT-SPOT device with 96 sample wells in a 12 x 8 well grid spaced at 1.0 cm intervals.
[0071] 100 μL of rabbit polyclonal IgG sample (1.0 μg) was added to each of the 8 wells in column 1.
[0072] In columns 2-12 are serial dilutions of antibody from 500ng-0.5ng, respectively.
[0073] Antibody-treated polyethylene sheets were dried overnight at 37°C.
[0074] The dried film was washed 3 times with pH 7.4 phosphate buffered saline (PBS).
[0075] Antigen was diluted to a final concentration of 1.0 μg / ml in tris buffer (TBS) containing 1% casein at pH 7.4.
[0076] 100 μL of antigen (representing 100 ng) was applied to...
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