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Immunofluorescence on Frozen Tissue Sections

JR Jason Reuter

Published: Dec 20, 2011 DOI: 10.21769/BioProtoc.163 Views: 30274

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Abstract

Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest. Immunofluorescence can also be used as a qualitative measure of protein expression.

Materials and Reagents

  1. Primary antibody
  2. Alexa fluor secondary antibodies (Life Technologies, InvitrogenTM)
  3. Horse serum (Santa Cruz Biotechnology, catalog number: sc-2483 )
  4. Hoechst 33342 nuclear dye (Life Technologies, InvitrogenTM, catalog number: H1399 )
  5. ProLong gold antifade mounting reagent (Life Technologies, InvitrogenTM, catalog number: P36934 )
  6. PAP pen (Sigma-Aldrich, catalog number: Z377821 )
  7. Nail polish (Thermo Fisher Scientific, catalog number: 50-949-071 )
  8. 100% ice cold acetone
  9. Phosphate buffered saline (PBS)
  10. Formaldehyde
  11. Methanol
  12. Triton X-100

Equipment

  1. -20 °C freezer
  2. Humidified chamber

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Immunology > Immune cell staining > Immunodetection

Biochemistry > Protein > Fluorescence

Biochemistry > Protein > Expression

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