Auto/Zyme Reagent Set
Most of the fixatives immobilize the tissue antigens by forming cross-linkages. Sometimes extensive cross-linking of proteins in high concentrations can also lead to the masking or destruction of antigens, which leads to poor staining or no staining at all. This loss of staining can be restored to certain extent by pretreatment of tissue sections with proteolytic enzymes. The unmasking of tissue sections is an important requirement for the optimal staining of antigens like immunoglobulins, Keratins, Carcinoembyonic antigen, Factor VIII-related antigen, Testosterone and to a lesser degree S-100 protein.
Auto/Zyme is a stabilized enzyme mixture designed for the uncovering of tissue antigens fixed with formalin.
Proteolytic enzymes are commonly used for the “unmasking” of tissue antigens in formalin-fixed, paraffin-embedded tissue sections. Groups of antigens such as immunoglobulins, keratins, carcinoembryonic antigen, factor VIII related antigen, testosterone, and to a lesser degree S100 protein, are optimally revealed after their pretreatment with proteolytic enzymes such as pronase, trypsin and pepsin. Auto/Zyme™ is a stabilized proteolytic enzyme mixture tested for its efficiency in the enhancement of the detection of these antigens. It is supplied as a two component kit: 1) enzyme concentrate and 2) diluting buffer. The final working solution is simply prepared by adding 2 drops of the enzyme concentrate to one mL of the diluting buffer. A total of 100 mL of the working solution can be prepared with the supplied reagents. Both of the supplied solutions are stable for six months when stored at 2-8°C.
Refrigerate at 4°C. Do not freeze.
Preparation of Working Reagent: Prepare this diluted reagent just before use and discard the unused portion after use.
1. Transfer 2 mL of Auto/Zyme diluent buffer into a test tube.
2. Add 4 to 8 drops of Auto/Zyme concentrate.* Mix reagents.
3. Apply this solution to tissue sections following their initial hydration step. Incubate for 5-10 minutes at 37°C.**
4. Wash tissue sections well with 1X Automation Buffer.***
* Amount required will depend on the length of fixxation of the tissue sections. 8 dops are recommended for sections subjected to longer fixations.
** This incubation time will vary dependeing on the degree of fixation of the tissue sections.
*** Biomeda's 10X Automation Buffer (Cat. No. M30).
Do not use beyond the expiration dated stated on the label.