XAF Black™ Lipofuscin Autofluorescence Blocker

Thaw the XAF Black™ Lipofuscin Autofluorescence Blocker DMSO solution to room temperature. If any precipitates are present, heat the vial at 60–70°C for 5 minutes before preparing the working solution.

1. Please follow a standard protocol to perform the following steps: tissue section fixation, deparaffinization, and antigen retrieval.

2. Add the XAF Black™ Lipofuscin Autofluorescence Blocker working solution and incubate at room temperature for 30 to 60 seconds.

3. Add mounting medium to the prepared sample, and then observe it under a fluorescence microscope.

1. To prepare the XAF Black™ Lipofuscin Autofluorescence Blocker working solution (1X), mix 50 μL of the XAF Black™ Lipofuscin Autofluorescence Blocker 20X DMSO stock solution with 1 mL of 70% ethanol.

   Note: Protect the working solution from light by either wrapping it in foil or storing it in a dark place.

   Note: For the best results, use this solution within a few hours of preparation.

   Note: Prepare 100 to 200 µL of 1X XAF Black™ Lipofuscin Autofluorescence Blocker working solution for each tissue section to be treated.

1. Follow your standard protocols to perform fixation, deparaffinization, and/or antigen retrieval on tissue sections.

2. Wash slides with PBS.

3. Remove PBS and place the tissue sections in a humidified slide chamber.

4. Add 100 to 200 µL of XAF Black™ Lipofuscin Autofluorescence Blocker working solution (1X) to each tissue sample, ensuring that the entire sample is fully covered.

5. Incubate the slides for 30 to 60 seconds.
   

   Note: The incubation time can be adjusted based on the requirements of the specific application to achieve optimal results.

6. Wash the slides with PBS twice.

7. Follow your recommended protocol to perform immunofluorescence staining using antibodies.

   Note: It is not recommended to use buffers containing detergents for blocking, antibody incubation, or washing steps. If detergents are necessary, please follow the post-treatment protocol.

8. Coverslip the slides using an aqueous-based fluorescence anti-fade mounting medium, such as FluoroQuest™ PLUS Antifade Mounting Medium (AAT Cat# 20008).

   Note: It is not recommended to use organic-based mounting mediums, as they are incompatible with the XAF Black™ Lipofuscin Autofluorescence Blocker.

1. Follow your standard protocols to perform fixation, deparaffinization, and/or antigen retrieval on tissue sections.

2. Follow your recommended protocol to perform immunofluorescence staining using antibodies.

   Note: Post-treatment with XAF Black™ Lipofuscin Autofluorescence Blocker may reduce fluorescence signals from antibodies or nuclear stains.

3. Wash slides with PBS.

4. Remove PBS and place the tissue sections in a humidified slide chamber.

5. Add 100 to 200 µL of XAF Black™ Lipofuscin Autofluorescence Blocker working solution (1X) to each tissue sample, ensuring that the entire sample is fully covered.

6. Incubate the slides for 30 to 60 seconds.
   

   Note: The incubation time can be adjusted based on the requirements of the specific application to achieve optimal results.

7. Wash the slides with PBS twice.

8. Coverslip the slides using an aqueous-based fluorescence anti-fade mounting medium, such as FluoroQuest™ PLUS Antifade Mounting Medium (AAT Cat# 20008).

   Note: It is not recommended to use organic-based mounting mediums, as they are incompatible with the XAF Black™ Lipofuscin Autofluorescence Blocker.