Sodium Citrate EDTA Antigen Retrieval Solution(40×)

When tissues or cells are fixed with aldehyde reagents such as paraformaldehyde or formaldehyde, cross-links are formed between proteins, which can obscure the antigen sites and weaken the staining signals during immunostaining. False positive staining results may also appear. To mitigate these issues, Sodium Citrate-EDTA Antigen Retrieval Solution(40×) uses sodium citrate buffer and EDTA to restore antigens by effectively removing the cross-links between proteins in samples such as paraffin sections. This greatly improves the immunostaining effect and is suitable for use with frozen sections and other samples as well. While frozen sections typically do not require antigen retrieval, it has been shown to still improve staining in some cases. Therefore, any sample fixed with aldehyde reagents may benefit from antigen retrieval to better expose the epitope and increase the effectiveness of subsequent immunostaining.

1. For paraffin sections:
(1) To remove the paraffin from the sections, a deparaffinization process is followed. Firstly, the sections are treated with xylene for two rounds of 5-minute intervals. Then, anhydrous ethanol is used for another two rounds of 5-minute intervals. Next, the sections are subjected to 90% ethanol for two rounds of 5-minute intervals. Afterwards, they are immersed in 70% ethanol for a duration of 5 minutes. Finally, the sections are rinsed twice for 5 minutes each in distilled water. This thorough deparaffinization process ensures the removal of paraffin from the sections.
To retrieve antigens, you will need to first submerge the slices in antigen retrieval solution (1×). Then, heat the solution at a temperature between 95-100° C for about 10-30 minutes. The appropriate heating time may vary depending on the sample and target proteins. Ensure that the antigen retrieval solution is preheated to the correct temperature before use. You can heat the solution using either a water bath or a microwave, but if you opt for the latter method, take care to avoid bumping the repair solution. Once heated, cool the solution to room temperature for approximately 20-30 minutes. Next, you will need to perform 1-2 washes with immunostaining washing solution, allowing each wash to last between 3-5 minutes. Once complete, you can proceed with subsequent immunostaining steps such as blocking.
2. Frozen sections: Wash the sections with immunostaining washing solution for 5 minutes. The process of antigen retrieval is like that of paraffin section.
3. Antigen retrieval of other samples can refer to the steps of paraffin section.

1. For your own safety, please protect yourself before using reagents, such as wearing lab coats and gloves.
2. Sodium citrate-EDTA antigen retrieval solution (40×) must be diluted 40 times with distilled water or MilliQ water before use to make antigen retrieval solution (1×).

Store at 2~8° C. If you do not use it for a long time, please store it at -20°C. The validity period is 12 months.

Citrate-EDTA Antigen Retrieval Solution(40×)