Transfected BHK-21 cells were collected following trypsinization (3–5 min) and centrifugation (300 × g, 5 min, RT), and total protein cell lysates were collected (10,000 × g, 10 min, RT) after homogenization with Tissue Extraction Reagent I (Thermo Fisher Scientific) Tris-based lysis buffer supplemented with Halt protease inhibitor cocktail (1X) (78,429, Thermo Fisher Scientific).
Halt protease inhibitor cocktail 1x
Manufactured by Thermo Fisher Scientific
Halt Protease Inhibitor Cocktail (1x) is a laboratory product designed to inhibit the activity of proteases, which are enzymes that break down proteins. It provides a convenient way to protect protein samples from degradation during extraction, purification, or other analytical procedures.
Automatically generated - may contain errors
3 protocols using halt protease inhibitor cocktail 1x
1
Protein Extraction from Transfected BHK-21 Cells
2
Postmortem Brain Tissue Preparation
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Postmortem brains and spinal cords were thawed and weighed to determine the mass in grams. Brains were bisected into forebrain and hindbrain pieces using a single cut with a scalpel blade between the striatum and hypothalamus. Tissue was homogenized in nine volumes of cold DPBS containing Halt Protease Inhibitor Cocktail (1x, Thermo Fisher Scientific) using a Precellys 24-bead beater (Bertin Instruments) with metal bead lysing matrix (MP Biomedical). Where necessary, brain lysates were clarified by centrifugation at 10,000 × g for 10 min at 4 °C. All tissue and samples were stored at −80 °C until further use.
3
Postmortem Brain and Spinal Cord Dissection
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Postmortem brains and spinal cords were thawed and weighed to determine the mass in grams.
Brains were bisected into forebrain and hindbrain pieces using a single cut with a scalpel blade between the striatum and hypothalamus. Tissue was homogenized in nine volumes of cold DPBS containing Halt Protease Inhibitor Cocktail (1x, Thermo Fisher Scientific) using a Precellys 24-bead beater (Bertin Instruments) with metal bead lysing matrix (MP Biomedical).
Where necessary, brain lysates were clarified by centrifugation at 10,000 × g for 10 min at 4°C.
All tissue and samples were stored at -80°C until further use.
Brains were bisected into forebrain and hindbrain pieces using a single cut with a scalpel blade between the striatum and hypothalamus. Tissue was homogenized in nine volumes of cold DPBS containing Halt Protease Inhibitor Cocktail (1x, Thermo Fisher Scientific) using a Precellys 24-bead beater (Bertin Instruments) with metal bead lysing matrix (MP Biomedical).
Where necessary, brain lysates were clarified by centrifugation at 10,000 × g for 10 min at 4°C.
All tissue and samples were stored at -80°C until further use.
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