Immunozyme-labeled antibody staining of cultured cells

Phytoenzyme histochemical technology uses enzyme as a marker for antigen-antibody reaction, and the enzyme catalyzes the corresponding substrate to form an insoluble reaction product. Light microscope observation requires that the final product formed is an insoluble colored precipitate, while electron microscope observation requires that the final product of the enzyme reaction has a higher electron density after proper treatment. Horseradish peroxidase (HRP) is currently the most widely used enzyme marker, with the advantages of strong stability and high specificity of enzyme reaction.

1) 4% paraformaldehyde prepared in 0.1mol / LPB (pH 7.2) was fixed in situ (4 â„ƒ) for 1 hour;

2) 2% H2O2 / methanol treatment (can be omitted), room temperature for 20 minutes, blocking endogenous peroxidase activity;

3) Incubate with 1% BSA for 15 minutes at room temperature after PBS rinse;

4) Incubate specific antibody (primary antibody) for 3 hours at room temperature or overnight at 4 â„ƒ;

5) After the PBS is fully rinsed, HRP or biotin-labeled secondary antibody is incubated at room temperature for 30-60 minutes. If it is a biotin-labeled secondary antibody, after the reaction, further operation according to the requirements of the ABC kit is required;

6) After PBS rinse, 1% glutaraldehyde 0.1m01 / LPB is prepared and fixed for 30-60 minutes, and PBS rinse;

7) Color reaction: 0.03% ï½ž 0.05% DAB 0.0l% H> (L / 0.05mol / LTrisâ€”HCl (pH 7.6) color development, stop the reaction in time, rinse with PBS;

8) 30% to 60 minutes after 1% OsO4;

9) Sample dehydration, in-situ embedding, ultra-thin section preparation and electron microscope observation are the same as conventional electron microscope.

Electron micrograph of cultured cells with immunoenzymatic antibody staining

Show thymus cell surface: thy1.2 antigen distribution (arrow)

Flight Case

Aluminum Flight Case,Aluminum Trolley Flight Case,Flight Case Plastic,Flight Case Parts

Changzhou Junye Aluminum Case Co., Ltd , https://www.junye-box.com