Basophil Degranulation Test Protocol

Basophils are involved not only in tachyphylactic hypersensitivity reactions but also in the body's anti-tumor immune response. Detecting the function of basophils helps in the diagnosis of allergic diseases and the prognosis of tumors. In this paper, a basophil degranulation test was performed to detect the function of basophils.

• Isolated basophils are washed twice with Hank's solution and centrifuged at 1500 r/min for 10 min each time.
• Aspirate the supernatant, leave about 0.3 ml of precipitate, and gently shake to resuspend the cells.
• Take 50 μl of allergen, mix it with 50 μl of cell suspension, place it in a 37°C water bath for 30 min, and replace the allergen with the same amount of buffer as a control tube.
• Add 150 μl of Alcian blue staining solution, mix well, and place at 15-25°C overnight. Count the number of basophils stained blue (F) in 9 large squares on the following day.

Creative Bioarray Relevant Recommendations

• Human basophils are a type of white blood cells. Basophils are the least common of the granulocytes, representing about 0.5 to 1% of circulating white blood cells. Creative Bioarray provides high-quality human basophils and their isolation kits.

• The whole process needs to be aseptic.
• The test subject must be identified. No fatty diet is allowed on the day of blood collection. Normal controls should have no history of allergy and should be collected and tested at the same time as the measurement group.
• There are many types of allergens, which should be selected rationally according to the different regions and seasons, combined with clinical information. Appropriate concentration should also be selected.

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