TRIS EDTA buffer

Product Composition: TRIS BASE and EDTA pH 9.00

Intended Use: Product for in vitro diagnostic use

Recommendations for Use:
Before use: The TRIS EDTA solution is 10x concentrated and therefore it must be diluted in a
1:10 range with distilled or deionized water (one part TRIS EDTA buffer and 9 parts water).
1. – Cut and mount sections on either charged or silane coated slides.
2A. – In case of using an automated PT Module for simultaneous dewaxing, rehydrating and antigen (HIER) retrieval, follow the instructions enclosed in the equipment data sheet. Basically, fill the tanks with 1.5 litres of diluted, ready-to-use buffer solution, place the slides without dewaxing into the racks and then immerse them into the buffer solution. Program the PT Module for a cycle of antigen retrieval of 20min at 95ºC and then cool to room temperature. Then follow with the step 4.
2B. – In case of not using the PT Module for HIER continue with dewaxing (using xylene or substitute) and rehydration of the tissue sections through decreasing series of ethanol. Then follow with the step 3 and 4.

3. – Unmasking: different alternative methods could be used to carry out the antigen retrieval process using the diluted TRIS EDTA buffer:
3.1 Unmasking in microwave: Place the sections in the diluted solution of TRIS EDTA (Coplin jar or other container with a volume as large as the laboratory considers) and bring the microwave oven to 700 watts (watt) for 10-15 minutes). Cool to room temperature.
3.2 Unmasking in pressure cooker: Fill the pressure cooker with 1-2 litres of diluted TRIS EDTA buffer. While hydrating the slides, heat the solution avoiding boiling. Then immerse the sections in the preheated buffer and close the pressure cooker. Wait until the pressure rises to the second notch and run from here 1-2 minutes (depending on the antigen unmasking). Remove and cool the pot (you can speed up the process placing the pot under running water). Open when there is no pressure and cool the slides inside of the pot for 20-30 minutes.
4. – Remove the sections and place them into the usual washing buffer (recommended TBS / TBS-Tween pH 7.5 – reference MAD-004077R)
5. – Proceed with the immunostaining protocol as usual

توضیحات و استفاده

Specificity, Interference and Limitation: TRIS EDTA buffer is used in methods for heat induced antigen retrieval (HIER). When used on material fixed in buffered formalin and embedded in paraffin, certain antibodies require antigen unmasking for retrieving the immunoreactivity that had been masked by the fixing process.

Storage: Store until the expiry date of the product at room temperature, away from any sources of intense heat or cold.

Application: The HIER has improved immunohistochemical staining in conventional surgical pathology as it improves the antigen unmasking and the staining intensity. The use of these techniques has its best advantages in case of nuclear antigens that presents a marked increase in signal intensity.

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