Enzyme markers.

: I want to know, how can enzyme labeling be very effective? What about the sodium periodide marker?
: Section 1 Indirect method testing
antibodies
basic principles
the specific
antigen
package is on the solid-phase carrier, forming a solid-phase antigen, added to the label to be tested This (containing corresponding antibodies), wherein antibodies and solid antigen antigens form antigen-antibody complex, and then add enzyme-labeled anti-antibodies (anti-human
immunoglobulin
antibodies, also known as secondary), and the above antigen-antibody complex binding. At this point, the substrate is added, and the enzymes on the complex catalyz the substrate to show color.
reagents
and equipment
purifying antigens.
antibody (spicy root peroxidase marker anti-
human immunoglobulin
)
substrate (phthalates OPD or tetamineTM
substrate buffer (Na2HPO4 0. 184g synthic acid 0.047g s/h2O10ml s/h2O2 5ul)
PBS (see Appendix)
pH9.6 carbonate buffer CBS (see Appendix))
lotion (PBS - 0.05% volume Tween20)
dilution (100ml PBS - 1g bovine
serum
blood protein)
termination fluid (2mol/L H2SO4)
ase label plate (polystyrene plate)
microplate
absorbent paper
seal tape
ase labeler
washboard machine, etc.
Experimental Step
I Preparation of an enzyme label plate for the test
(i) antigen package
1. Dilute the antigen into an appropriate concentration (reference: 5ug/ml) with a carbonate buffer, add the enzyme plate (50ul/hole) and seal it with tape overnight at 4 degrees C.
2. Discard the antigen solution, rinse 3 times with double steamed water,
naturally
and sealed with tape. This is a known antigen package by the enzyme plate, spare.
(ii) the concentration of the substrate groping
in the substrate buffer to add a different amount of substrate (reference: 5mg/10ml), add to the enzyme label plate (50ul/hole), seal with tape, 37 degrees C to avoid light to save 2h, in the absence of significant color change conditions to choose the largest possible base concentration.
(iii) the concentration of enzyme-labeled antibodies groping
dilute the enzyme-labeled antibody with dilution to an appropriate multiplus (reference: 40 to 4000 times), add the enzymatic label plate (50ul/hole) ensconced with antigens, sealed at 37 degrees C action 30min. Rinse with lotion 5 times in a row, then pat dry on absorbent paper, add the groped base seal at 37 degrees C action 2h, in the absence of color change conditions to choose the largest possible concentration of enzyme-labeled antibody.
.