10x Heat Mediated Antigen Retrieval Solution pH 6.0 (ab973)

Product Name 

10x Heat Mediated Antigen Retrieval Solution pH 6.0

Product type 

IHC reagents

Description 

10x Heat Mediated Antigen Retrieval Solution pH 6.0

Tested applications 
(see key)

IHC-P

Application notes 
(see key)

Recommended dilutions
Be sure to use heat resistant coplin jars

Procedure: Coplin Jar Method:

1. Dry tissue sections at 37°C for 1 hour and then dry slides 10-30 minutes at 60°C.
2. Fill the Coplin Jar with 50 ml of 1x Antigen retrieval solution. Fill a second Coplin Jar with 50 ml of deionized water (DI).
3. Place slides in the Coplin Jar.
4. Place both Coplin Jars in the pressure cooker chamber.
5. Place the lid on the chamber and start the heating process
6. When completed, open the chamber and allow solution to cool for 5 minutes.
7. Transfer slides with a forcep into the hot rinse (second Coplin Jar containing the DI water).
8. Continue cooling for 5 minutes.
9. Wash briefly in deionized water and rinse with PBS

Procedure: Tissue-Tek Staining Dishes:

1. Dry tissue sections at 37°C for 1 hour and then dry slides 10-30 minutes at 60°C.
2. Place slides in a slide holder and place in a Tissue-Tek staining dish.
3. Fill the staining dish with 250 ml of 1x Antigen retrieval solution. Fill a second staining dish with 250 ml of deionized water (DI).
4. Place both staining dishes in the pressure cooker chamber.
5. Place the lid on the chamber and start the heating process.
6. When completed, open the chamber and allow solution to cool for 5 minutes.
7. Transfer slides into the hot rinse (second Tissue-Tek staining dish containing the DI water).
8. Continue cooling for 5 minutes.
9. Wash briefly in tap water and rinse with PBS.


Important Technical Notes:

1. There are several heating methods that may be employed. These methods include: microwave oven, pressure cooker , hot water bath and steamer. For microwave use, licensing may be required under U.S. patent No. 5,244,787.
2. If using a pressure cooker, heat slides for 2-3 minutes and cool for 10 minutes.
3. Use positively charged slides and cut tissues 4-5 microns. Do not use any adhesives in the water bath. Poor fixation and processing of tissues will cause tissue sections to fall off the slides, especially fatty tissues such as breast. Tissues should be fixed a minimum of 6-12 hours.

Research areas 

Kits/ Lysates/ Other >> Tools and Reagents >> IHC Tools/ Reagents

Relevance 

This Antigen retrieval solution is a 10x buffer solution with paraffin emulsifier, allowing the user to deparaffinize and pretreat tissues in one-step. Xylenes and alcohols are not needed for deparaffinization, thus saving valuable technician time and reducing costs for reagents and hazardous waste. Antibody titers are doubled when compared to citrate buffer, pH 6.0 and background staining is reduced significantly. This product is specially formulated for superior pH stability at high temperature, and will help prevent the possibility of losing pH sensitive antigens.

Storage buffer 

PBS pH7.3 with protein carrier and preservative

Form 

This Antigen retrieval solution comes as a 10X solution.
The dilution is given as a ratio of concentrate volume to final volume.
Mix 1-part of the concentrated Antigen retrieval solution with 9 parts of deionized water (1:10).
This Antigen retrieval solution is non-toxic, non-flammable and is odorless
It is sodium azide and thimerosal free.

Storage instructions 

Store at room temperature.

Notes 

This Antigen retrieval solution is citrate buffer pH6. It incorporates ASSURE™, which is a color-coded pH indicator technology. The end-user is assured, by visual inspection, that the solution is at the correct dilution and pH. The 10x solution is orange. The 1X solution is yellow. When boiling, the solution becomes a pale orange. Should the pH rise above 7.0, the solution will turn a reddish color.

References for 10x Heat Mediated Antigen Retrieval Solution pH 6.0 (ab973)

  This product has been used in:

Iacobini C et al. Development of age-dependent glomerular lesions in galectin-3/AGE-receptor-3 knockout mice. Am J Physiol Renal Physiol 289:F611-21 (2005). PubMed: 15870382

If you publish research using ab973 please let us know so that we can cite the reference on this datasheet.

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