Nupage lds lithium dodecyl sulfate sample buffer

Manufactured by Thermo Fisher Scientific

The NuPAGE LDS (lithium dodecyl sulfate) sample buffer is a laboratory product used for sample preparation in protein analysis techniques such as SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). It is a denaturing buffer that helps solubilize and unfold proteins prior to electrophoretic separation.

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Lab products found in correlation

Biotin azide, by Thermo Fisher Scientific (1 mentions) 5 ethynyl 2 deoxyuridine (edu), by Thermo Fisher Scientific (1 mentions) Bioruptor, by Diagenode (1 mentions) Protease inhibitor, by Roche (1 mentions)

2 protocols using nupage lds lithium dodecyl sulfate sample buffer

Mammalian and Bacterial Cell Lysis Protocols

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Lysis buffer for mammalian cell lysis contained 50 mM Tris/HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 3% (w/v) CHAPS, 1 mM sodium orthovanadate, 10 mM sodium 2-glycerophosphate, 50 mM sodium fluoride, 10 mM sodium pyrophosphate, 0.27 M sucrose, 0.1% 2-mercaptoethanol, 1 mM benzamidine and 0.1 mM PMSF. Lysis buffer for E. coli protein purification contained 50 mM Tris/HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% (v/v) Triton X-100, 1 mM sodium orthovanadate, 10 mM sodium 2-glycerophosphate, 50 mM sodium fluoride, 10 mM sodium pyrophosphate, 0.27 M sucrose, 0.1% 2-mercaptoethanol, 1 mM benzamidine and 0.1 mM PMSF. Buffer A contained 50 mM Tris/HCl (pH 7.5) and 0.1 mM EGTA. TBST (TBS containing Tween 20) consisted of Tris/HCl (pH 7.5), 0.15 M NaCl and 0.2% Tween 20. SDS sample buffer was 1× NuPAGE LDS (lithium dodecyl sulfate) sample buffer (Invitrogen), containing 1% (v/v) 2-mercaptoethanol. Homogenization buffer for cellular fractionation contained 50 mM Tris/HCl (pH 7.5), 10 mM KCl, 1.5 mM MgCl₂, 0.1 mM EDTA, 0.5 mM EGTA and 44 mM sucrose. RIPA buffer contained 50 mM Tris/HCl (pH 7.5), 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% (v/v) Triton X-100, 0.1% 2-mercaptoethanol, 1 mM benzamidine and 0.1 mM PMSF.

vanderWijst J., Blanchard M.G., Woodroof H.I., Macartney T.J., Gourlay R., Hoenderop J.G., Bindels R.J, & Alessi D.R. (2014). Kinase and channel activity of TRPM6 are co-ordinated by a dimerization motif and pocket interaction. Biochemical Journal, 460(Pt 2), 165-175.

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Detection of Nascent and Stalled DNA Forks

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iPOND was essentially performed as described previously (Sirbu et al., 2011 (link)). In brief, 2 × 10⁸ cells were labelled with 10 μM EdU (Life Technologies) for 10 min to detect nascent forks. For stalled forks, cells were subsequently treated with HU for 2 h in the continued presence of EdU. Cells were crosslinked with 1% formaldehyde for 10 min at room temperature and quenched with 0.125 M glycine. For EdU conjugation with biotin azide, cells were permeabilized with 0.25% Triton X-100 in PBS buffer for 30 min, and then subjected to click–iT reaction buffer (10 mM sodium-L-ascorbate, 20 μM biotin azide [Life Technologies] and 2 mM CuSO4) for 2 h at room temperature. Cells were resuspended in lysis buffer (50 mM Tris-HCl (pH 8.0) and 1% SDS) supplemented with protease inhibitors (Roche), followed by sonication at 4°C using a Bioruptor (Diagenode). After centrifugation, EdU-labeled DNA was immunoprecipitated from supernatants by incubation with streptavidin-MyOne C1 beads (Life Technologies). Beads were washed and captured proteins were eluted by boiling beads in 2 × NuPAGE LDS (Lithium dodecyl sulfate) sample buffer (Invitrogen) containing 200 mM dithiothreitol for 35 min at 95 °C. Proteins were resolved by electrophoresis and d etected by western blot with the indicated antibodies.

Kim J., Sturgill D., Sebastian R., Khurana S., Tran A.D., Edwards G.B., Kruswick A., Burkett S., Hosogane E.K., Hannon W.W., Weyemi U., Bonner W.M., Luger K, & Oberdoerffer P. (2017). Replication stress shapes a protective chromatin environment across fragile genomic regions. Molecular cell, 69(1), 36-47.e7.

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