Teacher Meng Consulting: Appreciation of LDH in cell supernatant
On the first day after the festival, Heilongjiang Medical University inquired: I have some problems with the cell supernatant, which is a bit dazed, I hope the technology can answer the doubts. According to the description of Meng teacher. The LDH of the culture supernatant of mouse hepatoma cells was measured, and the LDH maximal release well (in the cell supernatant compared with 2% Triton) and the natural release well (cell supernatant was added to the empty medium) were designed. The maximum release hole LDH measurement value is lower than the natural release hole, and the cause is not understood.
Shanghai Jinma technical analysis:
tritonX100 is a surfactant. It has a hydrophobic end and a hydrophilic end, which can dissociate the membrane protein from the cell membrane, thereby achieving the purpose of extracting the membrane protein, but it does not maintain the enzyme activity. Excessive tritonX100 will destroy the surface hydrophilic layer of the protein and destroy the protein structure and reduce the enzyme activity. Triton X-100 can increase the permeability of eukaryotic cell membranes, but PBS or TBS containing 0.1%-0.5% Triton X-100 is often used. Whether the concentration used by the teacher is too high, leading to the loss of enzyme activity.
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