Rat Serum

For Immunohistochemistry (IHC) and Immunocytochemistry (ICC) 1. If tissues are used, deparafinize and hydrate tissue sections through xylene or other clearing agents and graded alcohols.(For frozen sections or cell smears; use unfixed, acetone fixed or appropriate fixative for the antigen in question; for cell smears it may be necessary to permealize the cell by detergent, please refer to antibody protocol. 2. Wash 2-3 with distilled or deionized water. 3. Incubate sections/cell smear in Endoblocker (#1) for 5-10 minutes at room temperature or 37°C. Note: If antigen retriever is required it can be applied after this stage. 4. Wash slide with PBS or Tris saline (with 0.02-0.05% nonionic detergent, Triton X100, Tween 20 or NP-40) or washing buffer (Immuno Automation buffer IBSC cat # AR-6561) 3-5X. 5. Incubate sections/ cell smear in Protein blocking solution (diluted animal serum), for 10 minutes. at RT or 37ºC 6. Wash slide with PBS 1X. 7. Follow instructions for IHC/ICC. The diluted serums are also used as blocking agents in all immunoassays.

This serum is used as blocking agents at 5% in buffer with all Immunoassays, to reduce non-specific background and conserved sequence. This serum can also be used as a negative control in immunoassays with proper dilution. It is recommended that the diluted serum should be used from the same host animal where the labeled antibody is produced; e.g. if the labeled antibody is made in rat the blocking serum should be diluted rat serum. This serum is obtained from pools of healthy adult animals. The serum is heat inactivated at 56ºC for 30 minutes and dilpidated and filtered. Each serum is tested with appropriate biotinylated antibody for cross-reactivity.