Antigen Demasker Reagent 4X
Antigen Demasker Reagent (Antigen Demasker is a new Antigen unmasking system formulated without the use of toxic heavy metal salts.)
Used as antigen demasker on formalin-fixed paraffin-embedded tissue sections for Immunohistochemistry.
PREPARATION: This reagent is supplied as 4X solution. Dilute as needed 4X with distilled or deionized water.
Many important cellular antigens are altered during tissue fixation. Particularly problematic are formalin-fixed specimens where the cross-linking effect of formalin is mainly responsible for the masking of these tissue antigens. However, many of these antigens can be unmasked and identified by immunocytochemical techniques with dramatic results when compared to untreated specimens. Using Biomeda's Antigen Demasker Reagent and a regular pressure cooker, a considerable increased in the identification of many otherwise difficult to detect antigens can be achieved. This simple 10 minute procedure not only permits many antigens to be detected, it also translates in considerable savings since it permits the further dilution of the primary antibodies employed in the detection. Additionally, the Antigen Demasker Reagent does not contain heavy metal pollutants and is safe and easy to use.
Refrigerate at 4°C or store at RT. Do not freeze.
Storage and Disposal: Store Antigen Demasker Reagent at 4-8°C or RT. Dilute 1:4 with distilled or deionized water. This diluted reagent can be reused for approximately 2 weeks. Since Antigen Demasker Reagent does not contain heavy metal pollutants, it can be disposed of according to local disposal regulations. Treating tissue sections with high temperature antigenic unmasking technique and antigen demasker reagent can produce irregular bleach in the staining pattern (See specification sheet of antibodies for detail). These can be identified by loss in the morphological detail and low level of hematoxylin counterstaining. Areas conforming to this bleached pattern should not be taken into consideration during the evaluation of the specimen's stain.
PROTOCOL FOR HIGH TEMP. ANTIGEN UNMASKING TECHNIQUE: 1. Deparaffinize and rehydrate tissue sections. Bring them to 1X Automation Buffer (Cat.No. M30) or PBS. 2. Place a pressure cooker on a heat source. Place slides in Copland jar or any steel staining rack with holder. Fill the jar or holder with antigen damasker 1X solution. The slides should be completely submerged in the solution. Cover and lock the pressure cooker. Continue heating on the heat source. Incubate tissue slides for 5 minutes after the pressure cooker begins to rock gently, run cold water on the pressure cooker. Do not open the lid until the air vent/cover lock drops. Open lid very carefully, take out tissue slides. PRESSURE COOKER IS SUPER HOT, BE VERY CAREFULL. Wash the slides 3 times in 1X Automation Buffer or PBS.
ALTERNATELY Slides can be boiled in antigen demasker solution 10-20 mins followed by cooling at RT for 10-20 mins. Transfer slides in Buffer. Proceed with immunostaining.