DTTdithiothreitolTFAtrifluoroacetic acidHigh molecular weight (HMW) subunits of glutenin, corresponding to those for which full length gene sequences are available, were purified and their M r s determined by mass spectrometry [matrix-assisted laser desorption ionisation (MALDI)]. In general, the M ...
The predicted molecular weight (MW) of the protein is the sum of the molecular weights of all protein amino acids. It is easy to calculate, e.g., using the freeonline ExPASy tool.Often the calculated MW is different from that observed on th...
Please note: 20% β-Mercaptoethanol (or 100 mM DTT) for the 4X SDS sample buffer might help to remove unspecific bands due to dissociation of the protein complex. Figure 7: NQO1 (11451-1-AP) is an enzyme that serves as a quinone reductas...
Cells were resuspended in Ni-Wash buffer (25 mM HEPES, 500 mM NaCl, 10% glycerol, 1 mM DTT, 10 μM DHT, 1% Tween-20, 20 mM imidazole at pH 7.4), lysed and centrifuged. Soluble protein was captured by IMAC and eluted with 500 mM imidazole. During an overnight ...
The resin was washed extensively with wash buffer containing 25 mM Hepes, pH 7.4, 150 mM NaCl, 2 mM DTT, and 0.005% (w/v) LMNG. The target hSMPD2 proteins were eluted with wash buffer plus 5 mM D-Desthiobiotin (IBA) and concentrated to a final volume of approximately 100...
WeightMarker(Low)Shippingat,20℃toredat,20℃odeNo.3450ize,for200lanesSuppliedReagent,5×LoadingBu,er1ml 1MDTT(Dithiothreitol)100μl LotNo. Concentration,12μg/μl Volume,50μl Form, 50mMTris-HCl,pH68 1mMEDTA 200mMNaCl 50,Glycerol Description, Thismarkerconsistsof6kindsofproteinswithaMolecularW...
HEPES, 0.5 mM EDTA) and treated for 5 min on ice with 10 μl of 10% NP-40 to remove the ciliary membrane. After centrifugation (10 min at 21,100×gat 4 °C), the axonemes were resuspended in a lysis buffer (50 mM Tris–HCl, pH 7.4, 500 mM NaCl, 0.4% SDS, 1 mM DTT),...
The denatured protein mixtures with blue loading dye and 30x DTT were loaded onto SDS-PAGE gels for electrophoresis and then blotted onto an Immobilon®-FL PVDF membrane (Millipore). After blocking with 5% skim milk, the membrane was incubated with the corresponding primary antibodies overnight,...
After Ni-NTA elution, the eluted protein was concentrated and dialyzed in reverse IMAC buffer (25 mM Tris pH, 50 mM NaCl, 1 mM DTT), and the His-tag was removed by TEV protease overnight at 4 °C. The protein pool was batch incubated with Ni-NTA resin, and untagged Wzt-...
All protein samples had a concentration of 10 µM in 10 mM Na-phosphate pH 7.0, 0.5 mM DTT and 0.1% LDAO. Isothermal titration calorimetry ITC experiments were performed with a MicroCal PEAQ-ITC instrument (Malvern Panalytical) at 25 °C in 10 mM HEPES pH 7.0, 20 mM ...