Antigen retrieval reagent

ImmunoSaver

ImmunoSaver is an antigen retrieval reagent for formalin-fixed tissues. This product is used to restore antigenicity that was reduced by formalin fixation. As a result, the quality of immunostaining is improved. While various conditions have to be examined in conventional methods, this product can retrieve a wide range of antigens under the same conditions.

Features

  • It restores the antigenicity of formalin-fixed tissues.
  • It improves the quality of immunostaining.
  • It retrieves a wide range of antigens under the same conditions.

Composition

10% sodium citraconate solution

Usage

  1. Dilute ImmunoSaver by a factor of 200 with distilled or deionized water, and heat to 98°C.
  2. Place deparaffinized tissue sections in the diluted solution, and incubate at 98°C for 45 minutes.
  3. Wash the tissue sections with PBS three times for 5 minutes each, and proceed to immunostaining.

Example of staining

  1. Treat formalin-fixed human pancreatic paraffin sections with ImmunoSaver.
  2. Wash the paraffin sections once with PBST (PBS containing Triton X-100 at a concentration of 0.2%) and twice with PBS.
  3. Block at room temperature for 1 hour.
  4. Rinse the sections gently with PBS and react them with the primary antibody diluted in PBST at 4°C overnight. (rabbit anti SNAP-25 1/8,000)
  5. Wash the sections once with PBST and twice with PBS.
  6. React the sections with the biotin-labeled secondary antibody at room temperature for 1 hour.
  7. Wash the sections once with PBST and twice with PBS.
  8. React the sections with peroxidase-labeled avidin (1/400, diluted in PBS) at room temperature for 1 hour.
  9. Wash the sections once with PBST and twice with PBS.
  10. Use DAB as substrate for chromogenic reaction.
  11. Wash the sections with distilled water for 5 minutes, three times.
  12. Stain the sections with Mayer's acidic hemalum solution for approximately 1 to 3 minutes.
  13. Wash the sections with running water, rinse with distilled water two or three times, and discriminate color with 70% ethanol.
  14. Clear and mount the sections.

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For research use or further manufacturing use only. Not for use in diagnostic procedures.

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