The effects of factors (extraction time, irradiation power, number of cycles) and their first order interactions were evaluated from antioxidant activity of extracts using the 2,2′-diphenyl-1-picrylhydrazil (DPPH) free radical scavenging method, the ferric reducing ability of plasma (FRAP) assay, and the estimation of total phenolic content using the Folin Ciocalteu method.
leaf solvents extracts using the broth micro dilution method recommended by the Clinical and Laboratory Standards Institute (CLSI) [18].
β-actin was detected in whole cell extracts using the AC-74 antibody (Sigma).
Cleaved NOTCH1 was detected in whole cell extracts using the antibody recognizing the V1744 epitope (Cell Signaling Technology).
In a separate experiment, Wnt3a was immunopurified from 35S-labeled Oto+Wnt3a insoluble extracts using the same method.
During our global analyses of a variety of cellular lipid extracts using the shotgun lipidomics technology we have recently developed [18], [19], an intriguing phenomenon has been observed.
Enzyme activity was measured in ARPE-19 cell extracts using the fluorogenic substrate 4-methylumbelliferyl-N-acetyl β-glucosoamide (Sigma) and previously published methods [23].
PLD activity was measured in 50 µg of cell extracts using the Amplex Red phospholipase D assay kit (Invitrogen Corporation, Carlsbad CA) according to the manufacturer's instructions.
Myotubes were harvested and the Unc45b was extracted, fractionated and affinity-purified from the cell extracts using the Flag epitope tag (Fig. 1).
Both p35 and Cdk5 were detected in the immunoprecipitates obtained from brain extracts using the anti-AATYK1 antibody (Fig. 1B, lane 3).
To further confirm these interactions, we performed GST pull-down experiments from HeLa mitochondrial extracts using the mature form of human frataxin (aa 81 210) fused to an N-terminal GST (GST-hFXN81 210). GST-hFXN81 210
