Immunofluorescence Protocol For Frozen Sections

Store frozen blocks at 80 ºc.

Immunofluorescence protocol for frozen sections.

Immunofluorescence on frozen sections. The suggested cryostat temperature is between 15 and 23 c. The section will curl if the specimen is too cold. Immunofluorescence staining protocol.

Microscope slides pre coated. Nagy gertsenstein vintersten and behringer ed. 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.

Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest. Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds. Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining. Mount tissue sections onto gelatin or poly l lysine coated slides by placing the cold sections onto warm slides.

Brigitte arduini version 1 2015 mar 23. Immunocytochemistry and immunofluorescence protocol related fluorescence. 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. Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry.

Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method. See cryoprotection and processing of embryonic tissue protocol. Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or. Slides can be safely stored for 6 12 months at 80 c until ready for staining.

Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1. Immunohistochemistry protocol for frozen sections. Cut cryostat sections at 5 10 µm and mount on gelatin coated histological slides. Protocol for immunofluorescent staining of mouse frozen sections tissue.

Immunofluorescence general protocol important. Immunofluorescence can also be used as a qualitative measure of protein expression. Preparation of slides. Do not allow frozen tissue to thaw before cutting.

Embed the tissue completely in oct compound prior to cryostat sectioning. Direct vs indirect if. Icc and if video protocol. 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.

Tissue preparation cyropreservation.