Antibody detection method
An antibody detection and antibody technology, applied in the biological field, can solve the problem of high equipment requirements
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Embodiment 1A
[0053] Example 1ACE2 coupled latex microspheres
[0054] 1) 390uL of cross-linking solution (50mM HEPES, pH 6.1), add 100uL of latex microspheres (JSR), shake and mix on a shaker to obtain a latex microsphere solution;
[0055] 2) Add 13uL of EDAC solution to the latex microsphere solution, shake and activate at 37 degrees Celsius for 1 hour;
[0056] 3) Add 50ul ACE2 / RBD (2mg / ml) to the latex microsphere liquid, and shake and cross-link at 37 degrees Celsius for 4 hours;
[0057] 4) Add 50ul of TW-20 solution, shake and seal at 37 degrees Celsius for 1 hour;
[0058] 5) Add the cross-linking liquid to the centrifuge tube, after centrifugation, carefully discard the supernatant with a pipette;
[0059] 6) Add 2 mL of washing solution (50 mM glycine + 0.1% NaN3, pH 8.0), vortex, centrifuge again, and carefully discard the supernatant;
[0060] 7) Add 5 mL of latex microsphere suspension (50 mM glycine+0.1% BSA+10% sucrose+0.1% NaN3, pH 8.0), and ultrasonically disperse.
Embodiment 2
[0061] Example 2 Standard preparation
[0062] Neutralizing antibody standards were diluted with matrix serum to the following concentrations: 0, 50ng / ml, 100ng / ml, 200ng / ml, 500ng / ml, 1000ng / ml, 2000ng / ml, 4000ng / ml, 8000ng / ml.
Embodiment 3
[0063] Example 3 Preparation of sample diluent
[0064] Both RBD polymer fragments and RBD monomer fragments were purchased from Guangdong Feipeng Bio, and the SDS-PAGE electrophoresis results are as follows figure 1 As shown, it can be determined from the molecular weight that lane 2 is the RBD multimer fragment.
[0065] 3.1 Use recombinant RBD antigen (RBD polyfragment) to prepare the following sample diluents with different concentrations. The basic formula of the diluent is: Tris buffer
[0066] Diluent 1 Diluent 2 Diluent 3 Antigen concentration 40ug / ml 25ug / ml 10ug / ml
[0067] 3.2 Use the recombinant RBD antigen (RBD monomer fragment) to prepare the following sample diluents with different concentrations. The basic formula of the diluent is: Tris buffer
[0068] Diluent 4 Diluent 5 Diluent 6 Antigen concentration 40ug / ml 25ug / ml 10ug / ml
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