Proteins in cell lysates were electrophoresed on SDS-polyacrylamide gels and transferred to Immobilon-P membranes (Millipore, Billerica, MA, USA). Membranes were preincubated with 5% skim milk in TTBS (tris-buffered saline containing 0.1% Tween 20) for 30 min and incubated overnight with ...
SDS-PAGE and immunoblotting Samples were separated on SDS-polyacrylamide gel and proteins were transferred onto a nitrocellulose membrane (GE healthcare) using a wet transfer system (BioRad). The membrane was blocked in PBS with 5% skimmed milk ...
Sodium dodecyl sulfate polyacrylamide gel electrophoresis SLE: Systemic lupus erythematosus SPE: Solid phase extraction TCA: Trichloroacetic acid TDP43: TAR DNA-binding protein 43 tRNA: Transfer RNA UHPLC: Ultra-high performance liquid chromatography UPLC : Ultra performance liquid chromatography...
Dysregulation of gut microbial bilirubin reduction affects serum bilirubin levels, which can have substantial health implications. In moderate concentrations, bilirubin serves as an important antioxidant with potential health benefits6,7. However, elevated serum bilirubin concentrations can become toxic, leadin...
Samples were run on 10 to 20% Tris-Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels followed by electrophoretic transfer to polyvinylidene-difluoride membrane (Millipore, Bedford, MA). Polyvinylidene-difluoride membranes were boiled in PBS for 5 minutes, followed by Western blotting ...
The translation products were analysed by one and two-dimensional polyacrylamide gel electrophoresis. It is shown that estradiol affects the translation activities of a limited number of mRNAs. Some of the effects observed, such as the stimulation of PRL mRNA and the relative inhibition of GH mRNA...
(50 mM Tris–HCl, 6 M urea, 30% glycerol, 2% SDS, 1% DTT), followed by equilibration in buffer II (50 mM Tris–HCl, 6 M urea, 30% glycerol, 2% SDS and 260 mM iodacetamide) for 10 min before loading them onto polyacrylamide gels (12% SDS-PAGE) for the second-dimensional ...
(Fig.5b). The increase in p21 with scrambled control ODN was likely a result of non-specific cellular stress and 600 nM ODNs are more toxic to cells than 400 nM, showing the overall lower proliferation rate; however, the levels of p21 (Fig.5a) correspond inversely with cell proliferation...
exchanged into 0.1% formic acid/30% acetonitrile. The eluates were again dried to a volume of 35–40 μl. Half of each eluate was diluted with Laemmli buffer with β-mercaptoethanol and applied to a Bio-Rad mini-Protean TGX 4–20% polyacrylamide gels and run at 200 V. Gels were ...
& Mann, M. Mass spectrometric sequencing of proteins from silver stained polyacrylamide gels. Anal. Chem. 68, 850–858 (1996). 56. Rao, M. N., Holkar, A. G. & Ayyangar, N. R. A novel transacylation method for the synthesis of a-N-acyl-b-lactones; application to (±)-diacetyl...