Summary: Immunofluorescence is a technique allowing the visualization of a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye such as fluorescein isothiocyanate (FITC). There are two major types of immunofluorescence staining methods: 1) direct immunofluorescence staining in which the primary antibody is labeled with fluorescence dye, and 2) indirect immunofluorescence staining in which a secondary antibody labeled with fluorochrome is used to recognize a primary antibody. Immunofluorescence staining can be performed on cells fixed on slides and tissue sections. Immunofluorescence stained samples are examined under a fluorescence microscope or confocal microscope.
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Protocols
- Cell Staining for Immunofluorescence Microscopy (BD Biosciences)
Protocol includes coverslip preparation for adherent and hon-adherent cells, paraformaldehyde fixation, methanol/acetone fixation, blocking, incubation with primary antibody, or more than one primary antibody, secondary antibody and preparation for microscopy.
http://www.bdbiosciences.com/support/resources/pro...
Added: Sun Aug 26 2012, Hits: 4874, Reviews: 0 Write review Cached - Preparation and Staining of Frozen Tissue Sections (BD Biosciences)
Protocol for preparing frozen sections standard immonuhistochemical staining including section preparation, sectioning and staining.
http://www.bdbiosciences.com/support/resources/pro...
Added: Sat Jul 27 2002, Hits: 3873, Reviews: 0 Write review Cached -
A Semi-Permenant Mounting Medium for Immunofluorescence Microscopy
(House Ear Institute)
A recipe for Moviol, an inexpensive mounting medium that is easy to prepare.
http://www.hei.org/research/aemi/moviol.htm
Added: Tue May 14 2002, Hits: 2295, Reviews: 0 Write review Cached -
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.
http://ygac.med.yale.edu/mtn/antibodycleanup.stm
Added: Tue May 14 2002, Hits: 910, Reviews: 0 Write review Cached -
C. Elegans Embryo Staining
(Dr. Ian Chin-Sang)
Staining protocol for C. Elegans embryos for immunofluorescence microscopy.
http://research.abl.es/methods/1422/
Added: Mon Feb 06 2012, Hits: 1092, Reviews: 0 Write review -
Double Immunofluorescence stain
(Giorgio Cattoretti, Columbia University)
Any type of tissue is suitable for this technique, as long as the antigenicity for your antigen(s) is preserved. Use cytocentrifuged cells or frozen or paraffin dewaxed sections
http://icg.cpmc.columbia.edu/cattoretti/Protocol/I...
Added: Fri Aug 30 2002, Hits: 4432, Reviews: 0 Write review Cached -
Fluorescence Mounting Medium (Antifade)
(Spector Lab)
http://www.cshl.org/labs/spector/fluorescence_medi...
Added: Tue May 14 2002, Hits: 2563, Reviews: 0 Write review Cached -
Fluorescence Staining of Cells
(Yu-li Wang's Lab, University of Massachusetts Medical School )
Staining with fluorescent phalloidin and immunofluorescence staining
http://ylwang.umassmed.edu/protocol/cfs/stain.htm
Added: Fri Oct 17 2003, Hits: 3170, Reviews: 0 Write review Cached -
Fluorochrome Absorption and Emission Spectra
(BD Pharmingen)
Provide some useful information.
http://www.bdbiosciences.com/pharmingen/protocols/...
Added: Sat Jul 27 2002, Hits: 1561, Reviews: 0 Write review Cached -
General Immunofluorescence Protocol
(Rosen Lab, Department of Molecular and Cellular Biology, Baylor College of Medicine)
http://public.bcm.tmc.edu/rosenlab/protocols/gener...
Added: Sat Aug 24 2002, Hits: 10519, Reviews: 0 Write review -
Immunfluoresence on Chamber Slides
(Bowtell Lab)
http://grimwade.biochem.unimelb.edu.au/bowtell/cel...
Added: Tue May 14 2002, Hits: 784, Reviews: 0 Write review Cached -
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...
http://ygac.med.yale.edu/mtn/HAimmunocytol.stm
Added: Tue May 14 2002, Hits: 753, Reviews: 0 Write review Cached -
Immunofluorescence (with Methanol)
(D. Amberg Lab)
This is the protocol to use with the Botstein anti-actin rabbit antibodies
http://www.upstate.edu/biochem/amberg/protocols/Me...
Added: Tue May 14 2002, Hits: 1581, Reviews: 0 Write review Cached -
Immunofluorescence (without methanol)
(D. Amberg)
http://www.upstate.edu/biochem/amberg/protocols/no...
Added: Tue May 14 2002, Hits: 1253, Reviews: 0 Write review Cached -
Immunofluorescence of Cells
(The Morimoto Lab, Northwestern University)
This protocol is used for staining cells grown in 2-well glass chamber.
http://www.biochem.northwestern.edu/morimoto/resea...
Added: Mon Mar 29 2004, Hits: 4985, Reviews: 0 Write review -
Immunofluorescence Technique
(Spector Lab)
protocol for immunofluorescence on cells
http://www.cshl.org/labs/spector/immunofluorescenc...
Added: Tue May 14 2002, Hits: 6267, Reviews: 0 Write review Cached -
In Situ Hybridization of mRNA and Immunofluorescence
(Corbett Lab)
Yeast protocol
http://www.biochem.emory.edu/labs/acorbe2/Yeast_FI...
Added: Tue May 14 2002, Hits: 1459, Reviews: 0 Write review -
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.
http://ygac.med.yale.edu/mtn/Immu_protocols.stm
Added: Tue May 14 2002, Hits: 807, Reviews: 0 Write review Cached -
Preparation of cytospin from single cell suspension
(Giorgio Cattoretti, Columbia University )
http://icg.cpmc.columbia.edu/cattoretti/Protocol/I...
Added: Fri Aug 30 2002, Hits: 1382, Reviews: 0 Write review Cached -
Preparing a cell block from cell lines / cell suspensions
(Giorgio Cattoretti, Columbia University )
http://icg.cpmc.columbia.edu/cattoretti/Protocol/I...
Added: Fri Aug 30 2002, Hits: 944, Reviews: 0 Write review Cached -
Progesterone receptor/BrdU double Immunofluorescence
(Rosen Lab, Department of Molecular and Cellular Biology, Baylor College of Medicine)
http://public.bcm.tmc.edu/rosenlab/protocols/PR_Br...
Added: Sat Aug 24 2002, Hits: 1870, Reviews: 0 Write review -
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.
http://www.ucsf.edu/vonzast/feeding.html
Added: Tue May 14 2002, Hits: 1143, Reviews: 0 Write review Cached -
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.
http://www.ucsf.edu/vonzast/permeab.html
Added: Tue May 14 2002, Hits: 958, Reviews: 0 Write review Cached -
Staining Nonfilamentous Actin with Gc-Globulin
(Yu-li Wang's Lab, University of Massachusetts Medical School )
Includes direct and indirect methods
http://ylwang.umassmed.edu/protocol/cfs/dbp.htm
Added: Fri Oct 17 2003, Hits: 464, Reviews: 0 Write review Cached -
Vital Staining of Chromosomes with Hoechst 33258
(Yu-li Wang's Lab, University of Massachusetts Medical School )
http://ylwang.umassmed.edu/protocol/cfs/hoechst.ht...
Added: Fri Oct 17 2003, Hits: 1037, Reviews: 0 Write review Cached