Immunohistochemistry Protocol (Frozen Sections)

This Immunohistochemistry protocol is for frozen sample sections.

Immunohistochemistry Protocol (Frozen Section)

This Immunohistochemistry protocol is for frozen sample sections.

Fixation and Sectioning

  1. Freeze and section tissue samples.
  2. Fix sections in Bouin’s solution, -20°C acetone, -20°C methanol, or formaldehyde for 2-30 minutes. (NOTE: Fixation time will depend on species, tissue origin and size of tissue sections).

Blocking and Primary Antibody Incubation

  1. Block slides with 10% serum from the species of the secondary antibody or 8% BSA.
  2. Incubate for 30 minutes to 1 hour at room temperature in a humidified chamber.
  3. Wash samples in PBS containing 2% BSA for 5 minutes.
  4. Incubate slides in a humidified chamber overnight with primary antibody solution (primary antibody in PBS containing 2% BSA).
  5. Wash three times with PBS containing 2% BSA for 5 minutes.

Secondary Antibody Incubation and Detection

  1. Incubate slides in a humidified chamber with secondary antibody solution (secondary antibody in PBS containing 2% BSA) according to manufacturer’s protocols.
  2. Wash three times with PBS containing 2% BSA for 5 minutes.
  3. Detect according to manufacturer’s protocols.

 

http://blog.immunoreagents.com/immunohistochemistry-protocol-frozen-section/

 

ImmunoReagents, Inc. is a leading global manufacturer of quality antibodies and reagents used in pharmaceutical research, life science research and in vitro diagnostics. Product offerings include a wide range of immunochemistry reagents such as purified immunoglobulinsprimary antibodies, and secondary antibodies covering a broad spectrum of immunoglobulins from various species. ImmunoReagents also provides custom manufacturing to meet specific customer requirements while adhering to cGMP guidelines and ISO quality systems requirements. The company is located in Raleigh, North Carolina