Technovit H7100 / H8100
EMS Catalog #14653 - 14654
Staining Protocols For Lymphoid Tissue
Normal immunostaining procedures for routine markers on sections obtained from lymphoid tissue.
Procedure
- Dry the sections for two hours at 37°C on a slide warmer. This must be done whether the slides have been stored at 4°C for some time or just collected.
- Pretreat the sections with trypsin at 37°C. Trypsin concentrations must be determined separately for each antibody. Use trypsin solution that has been preheated at 37°C for 30 minutes. Cover section with at least 100 micro liters of solution.
- Wash in PBS for 10 minutes at room temperature. Refresh the buffer four to five times.
- Preincubate in normal serum from the animal species in which the second antibody is raised for 30 minutes at 37°C. Only perform this step if a specific background is present using a particular antibody.
- Drip off excess serum and apply the first antibody in an appropriate concentration and incubate for two hours at 37°C.
- Wash in PBS for 10 minutes at room temperature. Refresh the buffer four to five times.
- Block endogenous preoxidase in a solution of 0.05% hydrogen peroxide in phosphate buffered saline, pH 7.4, for 30 minutes at room temperature.
- Wash in PBS for 10 minutes at room temperature. Refresh the buffer four to five times.
- Incubate in appropriate dilutions of the secondary antibody, containing 5% normal serum for 60 minutes at room temperature.
- Wash in PBS for 10 minutes at room temperature. Refresh the buffer four to five times.
- Develop the peroxidase activity in daminobenzidine ( DAB ).
- Counterstain the sections in Hematoxylin or if a more advanced morphological detail is necessary, in periodic-acid-Schiff reagent.
- Cover with Glycerin-gelatin and cover glass.
Product Information
Technovit Glycol Methacylate Kits H7100 / H8100