Cut cryostat sections at 5 10 µm and mount on gelatin coated histological slides.
Immunofluorescence protocol for frozen sections.
Mount tissue sections onto gelatin or poly l lysine coated slides by placing the cold sections onto warm slides.
Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method.
Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or.
Preparation of slides.
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.
Microscope slides pre coated.
Modified from manipulating the mouse embryo 3.
The suggested cryostat temperature is between 15 and 23 c.
Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest.
Do not allow frozen tissue to thaw before cutting.
Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1.
Icc and if video protocol.
This protocol is also suitable for 40µm free floating.
Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry.
Immunofluorescence staining protocol.
Store frozen blocks at 80 ºc.
Embed the tissue completely in oct compound prior to cryostat sectioning.
See cryoprotection and processing of embryonic tissue protocol.
Brigitte arduini version 1 2015 mar 23.
Immunofluorescence on frozen sections.
Nagy gertsenstein vintersten and behringer ed.
The section will curl if the specimen is too cold.
Immunocytochemistry and immunofluorescence protocol related fluorescence.
Immunofluorescence can also be used as a qualitative measure of protein expression.
Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds.
Protocol for immunofluorescent staining of mouse frozen sections tissue.
Immunofluorescence on frozen tissue sections bio protocol.
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.
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.
Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining.
Immunohistochemistry protocol for frozen sections.
Immunofluorescence general protocol important.
Slides can be safely stored for 6 12 months at 80 c until ready for staining.
