GZMA/Granzyme A Protein, Human (HEK293, His), product number HY-P76377, is supplied as a lyophilized powder. I plan to reconstitute GZMA to a concentration of 100 μg/ml. What are the reconstitution steps?
I have purchased Lys-C (HY-P3205) from MedChemExpress, which is provided as a liquid solution. According to the instructions, it is prepared in 20 mM Tris HCl, pH 8.0. The recommended activation buffer for GZMA is significantly more alkaline (pH 9.0). What are the steps to prepare an activation buffer containing 0.2 µg/mL Lys-C using the HY-P3205 product?
Lily Watson
Add 100 µL of the supplied reconstitution buffer (ddH₂O). Then add 400 µL of carrier protein-containing buffer or culture medium (recommended options: 0.1% BSA, 5% HSA, 10% FBS, or 5% trehalose). → Final concentration should be 100 µg/mL (i.e., 100 ng/µL). It is recommended to use 0.1 M Tris, pH 9.0 activation buffer containing carrier protein, with a total volume of 500 µL. Aliquot into 25 µL portions per tube. Store at –20°C to –80°C for 3 to 6 months. Prepare 0.1 M Tris, pH 9.0 (referred to as the “activation buffer”). Dissolve Lysyl endopeptidase (Lys-C) in this activation buffer to prepare a working solution at 0.2 µg/mL. Mix the Lys-C activation buffer solution (0.2 µg/mL) with 100 µg/mL GZMA at a 1:1 volume ratio → resulting concentrations after mixing: GZMA 50 µg/mL, Lys-C 0.1 µg/mL. Incubate at 37°C for 1 hour to complete activation.
Reconstitution Procedure:
Activation Procedure:
The answer to this question is provided by MedChemExpress Technical Support. [email protected]
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