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