Immunohistochemistry protocol pdf

Wash the slides in ml PBS for 2 changes 5 min each. Wash slides in ml PBS for 2 changes, 5 min each. Allow the color development for less than 5 min until the desired color intensity is reached.

Caution : DAB is a suspected carcinogen. Handle with care. Wear gloves, lab coat and eye protection. Wash slides in ml PBS for 3 changes 2 min each. Optional: Counterstain slides by immersing sides in Hematoxylin e. Gill's Hematoxylin for min. Rinse the slides in running tap water for more than 15 min.

Clear the tissue slides in 3 changes of xylene and coverslip using mounting solution e. The mounted slides can be stored at room temperature permanently. Observe the color of the antibody staining in the tissue sections under microscopy. In most cases, this is true. Antigen retrieval helps both the accessibility of the antibody to the tissue and also counteracts the fixation effects on the recognized epitopes. Can antibody X be used for immunohistochemistry?

What concentration do I use? While we do not test for IHC application in house, we will indicate on the datasheet if an antibody has been published for use in this application. Tips: For initial experiments, the user must titrate primary and secondary reagents so that staining with the secondary antibody alone yields no background while staining with primary and secondary antibodies yields strong, specific staining.

Save Close Clear. Search X. Lab Timer X. Rinse the tissue with running tap water for 5 min. Cleared the tissue in xylene for 2 times, 5 min each. Immerse the tissue in paraffin for 3 times, 5 min each. Immunostain formalin-fixed, paraffin-embedded tissue sections Step : 9.

Deparaffinize slides in xylene for 2 times, 5 min each. Rinse with PBS for 2 times, 5 min each. Rinse slides with PBS for 2 times, 5 min each. Drain off the blocking buffer from the slides. Wash the slides with PBS for 2 times, 5 min each. Wash slides with PBS for 2 times, 5 min each. Wash slides with PBS for 3 times, 2 min each.

Optional Counterstain slides by immersing sides in Hematoxylin for min. Rinse the slides in running tap water for 10 min. Observe the color of the antibody staining in the tissue sections under microscopy.