免疫荧光

• Immunofluorescence Technique (Spector Lab)
  protocol for immunofluorescence on cells
    

• Immunofluorescence Protocol (Walter Steffen)

  • Methanol fixation

  • Formaldehyde fixation

  • Paraformaldehyde/glutaraldehyde fixation

  • EGS fixation

  • Fixation of the cytoskeleton

  • Preparation of cover glasses

  • Cell culture

  • Fixation protocol

  • Immunofluorescence protocol

  • Double immunofluorescence

  • Protein A - protein G

  • Avidin - biotin
      

• Immunofluorescence (Corbett Lab)
  Yeast protocol
    

• Immunfluoresence on Chamber Slides (Bowtell Lab)
    

• Immunofluorescence (with Methanol) (D. Amberg Lab)
  This is the protocol to use with the Botstein anti-actin rabbit antibodies
    

• Immunofluorescence (without methanol) (D. Amberg)
   

• A Semi-Permenant Mounting Medium for Immunofluorescence Microscopy (House Ear Institute)
  A recipe for Moviol, an inexpensive mounting medium that is easy to prepare.
    

• In Situ Hybridization of mRNA and Immunofluorescence (Corbett Lab)
  Yeast protocol
      

• Immunofluorescence Staining and Flow Cytometry of Intracellular Cytokines (eBioscience)
  A modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surface molecules and intracellular cytokines at single-cell level.  In this protocol, cells are first activated in vitro, stained for surface antigens, as in the regular staining protocol, then fixed and permeabilized to allow for anti-cytokine antibodies to stain intracellularly. 
     

• Fluorescence Mounting Medium (Antifade) (Spector Lab)
    

• Large-scale Immunofluorescence (YGAC)
  This protocol describes method for preparing cells for immunofluorescence, in which all incubations and washes are performed in microtiter dishes. Two dishes can be processed simultaneously. The last step is to transfer each sample to a well on a polylysine coated 8-well slide for visualization. The protocol can also easily be adapted for preparing cells for immunofluorescence in microfuge tubes. 
      

• Immunocytology using the HA Epitope(YGAC)
  The multiple copies of the HA epitope present in the HAT tag can be detected by the mouse monoclonal antibodies 12CA5 (Boehringer) and 16B12...  
     

• Staining for surface receptors
  by feeding antibody (Von Zastrow lab)
  This protocols allows the use of fluorescence microscopy to visualize epitope-tagged receptors expressed by stable adherent cell lines. In contrast to the permeabalized cell protocol, this protocol where antibody is "fed" to live unpermeabalized cells will only visualize receptors that started out on the surface of the cell.
    

·         Staining for total receptor(Von Zastrow lab)
This protocol will visualize both internalized receptors that were once on the plasma membrane and receptors in the biosynthetic pathway. This protocol was specifically written for visualizing agonist-induced internalization of receptors.

·         Antibody Cleanup(YGAC)
In order to decrease the amount of nonspecific staining, it is often necessary to preabsorb primary and secondary antibodies to yeast cells lacking the antigen prior to use.