Preparation method of common immune reagents

Preparation method of common immune reagents
1. Antibody or immunoglobulin dilution 0.01mol / L pH7.2PBS
NaH2PO4.2H2O 0.39g
Na2HPO4 1.27g
NaCl 0.85g
NaN3 200mg
H2O (distilled water) to 1000ml

2. DAB solution preparation: (histochemical and Western blot color development)
Weigh DAB 30mg, dissolve it with 0.05mol / L pH7.6Trs-HCl buffer 100ml, if there is precipitation, use it after filtration, generally fresh preparation.
Add 1% H2O2 1 ～ 1.5ml before use, H202 concentration is 0.01%, mix and use.

3.3,3,5,5, -Tetramethylbenzidine (TMB) solution preparation:
Use dimethylformamide (DMF) or absolute ethanol to prepare TMB to a concentration of 1% and store at 4 ℃ for half a year before use.
Before use, add 0.1ml (1mg TMB) 1% TMB solution with pH5.0 phosphoric acid-citric acid substrate solution 9.9ml, then add 1μl 30% H2O2 per ml TMB, mix well and use immediately.
After 2N H2SO4 was terminated, the OD value was measured at 450 nm.

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