We are trying to achieve better results in our immunohistochemistry assays (paraffin-embedded tissue sections) for antigen retrieval of surface markers such as CD3. Our current protocol of sodium citrate/citric acid buffer + heating seems to be too aggressive for the CD molecules or maybe it impairs the antibody affinity to the target.
Hi,
based on IHC world website and my experience Sodium citrate pH=6 is the best you should place your slide in the buffer container and then put in microwave for less than 2min until you see start bubbling then transfer it to steamer for 10 min and after 10min take them out let them cool for 10min then you can proceed to next step.
Good luck
check this. I have mention the name of wash buffer and stain buffer and blocking buffer. You can also used 10% goat serum for blocking purpose. I have used the below mention product, with sufficient reproducible result.
https://www.thermofisher.com/order/catalog/product/00-4954-56?ICID=search-product
http://www.bdbiosciences.com/us/applications/research/t-cell-immunology/th-1-cells/intracellular-markers/cytokines-and-chemokines/buffers/stain-buffer-fbs/p/554656
https://www.thermofisher.com/order/catalog/product/00-4952-54
For the CD-4 and the CD-8 I use an TRIS-EDTA buffer pH=9.
It gives better results than the citratebuffer although the morphology can be a little bit less.
10 X conc TRIS-EDTA buffer : 24 grams TRIS and 7.4 grams EDTA disodiumsalt dissolved in 2 litres of aqua destillata. (dissolve for at least 1 hour the pH will be 9.0)
before staining dilute the buffer 10 times.
succes Enno
- Badges
- Science method