Immunofluorescence Protocol For Frozen Sections

Cut cryostat sections at 5 10 µm and mount on gelatin coated histological slides.

Immunofluorescence protocol for frozen sections.

Icc and if video protocol. Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds. Embed the tissue completely in oct compound prior to cryostat sectioning. Modified from manipulating the mouse embryo 3.

Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry. This protocol is also suitable for 40µm free floating. 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. Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest.

Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1. Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining. Protocol for immunofluorescent staining of mouse frozen sections tissue. 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.

The suggested cryostat temperature is between 15 and 23 c. 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. Store frozen blocks at 80 ºc.

The section will curl if the specimen is too cold. Slides can be safely stored for 6 12 months at 80 c until ready for staining. Preparation of slides. Immunofluorescence on frozen tissue sections bio protocol.

Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or. This portion of the protocol can be skipped if you are working with pre mounted tissue slides. Immunohistochemistry protocol for frozen sections. Mount tissue sections onto gelatin or poly l lysine coated slides by placing the cold sections onto warm slides.

Do not allow frozen tissue to thaw before cutting. Immunocytochemistry and immunofluorescence protocol related fluorescence. Immunofluorescence can also be used as a qualitative measure of protein expression. Nagy gertsenstein vintersten and behringer ed.

Direct vs indirect if. See cryoprotection and processing of embryonic tissue protocol. Immunofluorescence on frozen sections. Immunofluorescence staining protocol.

Tissue preparation cyropreservation.