Immunofluorescence Protocol For Frozen Sections

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.

Immunofluorescence protocol for frozen sections.

Embed the tissue completely in oct compound prior to cryostat sectioning. Protocol for immunofluorescent staining of mouse frozen sections tissue. This protocol is also suitable for 40µm free floating. Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or.

Immunofluorescence staining protocol. Immunofluorescence general protocol important. Brigitte arduini version 1 2015 mar 23. Slides can be safely stored for 6 12 months at 80 c until ready for staining.

Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry. Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining. Preparation of slides. Microscope slides pre coated.

Immunofluorescence on 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. Store frozen blocks at 80 ºc. This portion of the protocol can be skipped if you are working with pre mounted tissue slides.

Immunohistochemistry protocol for frozen sections. Cut cryostat sections at 5 10 µm and mount on gelatin coated histological slides. Immunofluorescence can also be used as a qualitative measure of protein expression. Nagy gertsenstein vintersten and behringer ed.

Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1. 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. See cryoprotection and processing of embryonic tissue protocol. Direct vs indirect if.

Tissue preparation cyropreservation. The section will curl if the specimen is too cold. Immunocytochemistry and immunofluorescence protocol related fluorescence. Do not allow frozen tissue to thaw before cutting.

Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method. Icc and if video protocol. Modified from manipulating the mouse embryo 3. The suggested cryostat temperature is between 15 and 23 c.

Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds.