Immunocytochemistry and immunofluorescence protocol related fluorescence.
Immunofluorescence protocol for frozen sections.
The section will curl if the specimen is too cold.
Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1.
Cut cryostat sections at 5 10 µm and mount on gelatin coated histological slides.
Immunofluorescence on frozen tissue sections bio protocol.
Preparation of slides.
Store frozen blocks at 80 ºc.
Do not allow frozen tissue to thaw before cutting.
Please refer to the applications section on the front page of product datasheet or product webpage to determine if this product is validated and approved for use on cultured cell lines if ic paraffin embedded samples if p or frozen tissue sections if f.
Embed the tissue completely in oct compound prior to cryostat sectioning.
See cryoprotection and processing of embryonic tissue protocol.
Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method.
Icc and if video protocol.
This protocol is also suitable for 40µm free floating.
Microscope slides pre coated.
Immunofluorescence can also be used as a qualitative measure of protein expression.
Brigitte arduini version 1 2015 mar 23.
Tissue preparation cyropreservation.
Mount tissue sections onto gelatin or poly l lysine coated slides by placing the cold sections onto warm slides.
Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or.
The following is a general procedure guide for preparation and staining of acetone fixed frozen tissues using a purified unconjugated primary antibody biotinylated secondary antibody and streptavidin horseradish peroxidase sav hrp and dab detection system.
This portion of the protocol can be skipped if you are working with pre mounted tissue slides.
Paraffin and frozen sections reagents can be applied manually by pipette or this protocol can be adapted for automated and semi automated systems if these are available.
Protocol for immunofluorescent staining of mouse frozen sections tissue.
Nagy gertsenstein vintersten and behringer ed.
Immunofluorescence staining protocol.
The suggested cryostat temperature is between 15 and 23 c.
Modified from manipulating the mouse embryo 3.
Slides can be safely stored for 6 12 months at 80 c until ready for staining.
Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry.
Direct vs indirect if.
Immunofluorescence general protocol important.
Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest.
Immunofluorescence on frozen sections.
