100x Citrate Buffer pH 6.0 (ab64236)

Thank you very much for contacting us.

I have received a reply from a colleague who run the test in lab, for checking the change in pH when this product is diluted. They have confirmed that, with the dilution there was only a fraction of change in pH which was ˜0.4 from 7.98 to 7.56.

We are convinced that product we have sent is of high quality. There should not be a change in pH when diluted. Please check the diluents you have been using or retest the pH.

I would suggest using the product in Immunohistochemistry as recommended; this will work. In any case if this product does not work please contact me we will replace the product.

I hope this information will be helpful. Should you have any other question please do not hesitate to ask.

Thank you for contacting us.

The basic protocol for using a microwave with this buffer is as follows:

1. Deparaffinize and rehydrate tissue sections.
2. Dilute the product 1:100 with deionized water. Example: 2 ml of concentrate diluted in 198 ml of deionized water.
3. Immerse sections in Coplin jar filled with diluted Citrate Buffer reagent..
4. Microwave on HIGH power until boiling.
5. Keep the slides warm by heating for 10 minutes on LOW power.
6. Let Coplin jar sit in microwave for at least 20 minutes.
7. Remove and rinse with water..
8. Rinse with buffer and proceed with the appropriate staining protocol for the primary antibody in use.

You can subsitute any plastic rack and container for the Coplin jar. The main thing to avoid is allowing the sections to dry out, so make sure the sections are completely covered by the buffer at all times during and after the hot incubation.

You may want to test heating the buffer in the container without any slides, as it is very easy for the buffer to boil over (that is, to boil too much and too vigorously) so that some of the buffer is lost, exposing the sections to air. If using the microwave, it is important to control the boiling by using low power and watching the solution as it incubates, if necessary.

To avoid the boiling over that can occur in a microwave, a waterbath can be substituted for the microwave by simply heating the waterbath to 95-100oC and placing the container with the rack of slides and pre-heated buffer (or the Coplin jar with pre-heated buffer) in the hot waterbath for 15 - 20 minutes. The amount of time in the microwave or waterbath may need to be optimized but 15 - 20 minutes is usually sufficient, followed by 20 minutes to allow the buffer to cool, with the slides still in the container or Coplin jar.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.