Exact(6)
Anhydrous KBr was used as a pellet material.
The IB pellet material collected from the large scale production (700 ml of culture) was predominantly used for VEGF165 solubilisation and refolding experiments.
(5) Gas supply system, it can provide the pellet material of D2 or H2 and propellant gas of helium, and (6) control and diagnostic system.
Right: mitochondria were exposed to alkaline treatment at pH 12. Soluble (S) and membrane integrated (P, pellet) material was separated by centrifugation and analyzed by immunoblotting.
Two washes were then performed on the pellet material using 10 mM NaH2PO4 (pH6.5) and 1% Triton-X100, to remove any endoplasmic reticulum membranes.
Similar to CCMV CP, EM analysis from resuspended pellet material indeed confirmed the formation and presence of VLPs for CCMV CP-chimeric proteins containing a single 6xHis-tag at its N- or C-terminus and also when present at both termini simultaneously (Fig. 4b1-6).
Similar(54)
Insoluble extracts for gels only were prepared by solubilizing pelleted material in 0.5% SDS, 1% β-mercaptoethanol.
Insoluble extracts for immunoprecipitation (IP) were prepared by resuspending pelleted material in a small volume of 1% SDS, 20 mM Hepes pH 7, 150 mM NaCl, 10 mM DTT, heating for 10 min at 80°C, diluting 10× with SDS-free RIPA, then spinning at max speed for 10 min to pellet debris, and collecting supers in fresh tubes.
Pelleted material at the bottom of the gradient was also tested by western blotting.
Briefly, 2 mL of RNAprotect Bacteria reagent was added to cell pellets and gently vortexed to resuspend pelleted material.
Pelleted material was extracted in 1/10 original volume SDS-PAGE buffer to concentrate the final IP fraction.
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