Immunofluorescence Protocol For Frozen Sections

Immunofluorescence on frozen tissue sections bio protocol.

Immunofluorescence protocol for frozen sections.

Modified from manipulating the mouse embryo 3. Preparation of slides. The suggested cryostat temperature is between 15 and 23 c. Icc and if video protocol.

Slides can be safely stored for 6 12 months at 80 c until ready for staining. Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds. Brigitte arduini version 1 2015 mar 23. Immunofluorescence staining protocol.

Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method. See cryoprotection and processing of embryonic tissue protocol. Cut cryostat sections at 5 10 µm and mount on gelatin coated histological slides. Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry.

Nagy gertsenstein vintersten and behringer ed. Mount tissue sections onto gelatin or poly l lysine coated slides by placing the cold sections onto warm slides. Direct vs indirect if. Microscope slides pre coated.

This portion of the protocol can be skipped if you are working with pre mounted tissue slides. The section will curl if the specimen is too cold. Immunofluorescence general protocol important. Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest.

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. Immunofluorescence can also be used as a qualitative measure of protein expression. Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1. This protocol is also suitable for 40µm free floating.

Store frozen blocks at 80 ºc. Tissue preparation cyropreservation. 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. 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. Protocol for immunofluorescent staining of mouse frozen sections tissue. Immunofluorescence on frozen sections. Do not allow frozen tissue to thaw before cutting.

Immunohistochemistry protocol for frozen sections.