Sodium dodecyl sulfate (sds)

Manufactured by MP Biomedicals

Sourced in United States

SDS is a laboratory reagent used in biochemical applications. It is a detergent that aids in the solubilization and denaturation of proteins, facilitating their analysis and separation.

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Lab products found in correlation

7 protocols using sodium dodecyl sulfate (sds)

Polyelectrolyte Complex Formation

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All experiments were conducted using Millipore Direct-Q 3 deionized water with an 18.2 MΩ cm minimum resistivity. PAH (nominal molecular weight = 120–200 kDa) was purchased from Alpha Aesar (Ward Hill, MA). Ibuprofen and TPP were obtained from Sigma-Aldrich (St. Louis, MO), while HCl, NaOH and PBS powder were purchased from Fisher Scientific (Fair Lawn, NJ). The SDS and SDBS were obtained from MP Biomedicals (Solon, OH) and TCI America (Portland, OR), respectively. All materials were used as received.

de Silva U.K., Brown J.L, & Lapitsky Y. (2018). Poly(allylamine)/tripolyphosphate coacervates enable high loading and multiple-month release of weakly amphiphilic anionic drugs: an in vitro study with ibuprofen. RSC Advances, 8(35), 19409-19419.

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Glucose Uptake Assay in 3T3-L1 Adipocytes

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The 3T3-L1 fibroblasts were seeded on coated Poly-L-lysine (PLL) (Sigma-Aldrich) dishes and differentiated. Mature 3T3-L1 adipocytes were washed twice with warm Krebs–Ringer–phosphate–HEPES (KRPH) buffer (20 mmol/L HEPES, 5 mmol/L KH₂PO₄, 1 mmol/L CaCl₂, 136 mmol/L NaCl, and 4.7 mmol/L KCl set at pH 7.4 with NaOH) containing either 1 or 0 mmol/L Mg²⁺. Cells were incubated without or with 10 nmol/L human insulin (Sigma-Aldrich) and 0 or 1 mmol/L Mg²⁺ for 0, 10, 20 or 30 minutes at 37°C in radioactive buffer containing 5 mmol/L 2-deoxyglucose (2DG) (Sigma-Aldrich) and 1 μCi ³H-2DG (PerkinElmer, Hoogvliet Rotterdam, the Netherlands). The incubation was stopped by washing with cold KRPH buffer and cells were lysed with 0.05% (w/v) SDS (MP Biomedicals) in dH₂O. Cell lysates were added to Opti-Fluor O scintillation liquid (PerkinElmer) and counted using the Hidex 600 SL. Radioactive counts were corrected for the background count in each of the incubation buffers. The assay was repeated without the addition of ³H-2DG to correct for protein concentrations using the bicinchoninic acid (BCA) assay (Thermo Fisher Scientific) according to the manufacturer’s protocol.

Oost L.J., Kurstjens S., Ma C., Hoenderop J.G., Tack C.J, & de Baaij J.H. (2022). Magnesium increases insulin-dependent glucose uptake in adipocytes. Frontiers in Endocrinology, 13, 986616.

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Molecular Biology Technique Reagents

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Molecular-grade Tris, SDS, glycine, Coomassie brilliant blue R250, ethanol, and methanol were bought from MP Biomedicals, and polyvinylidene fluoride (PVDF) membranes from Millipore (MA, USA). Fmoc-amino acid trityl chloride resins, Fmoc-L-amino acid, coupling chemicals, and 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate were bought from Chempep (Miami, FL). Na₂EDTA, skim milk, FBS, MTT, NaCl, KHPO₄, KCl, and NaHPO₄ were bought from Himedia (India). The chemicals such as agarose, Trizma base, magnesium acetate, acetic acid, EDTA, ethidium bromide and 6× loading dye were purchased from Sigma-Aldrich. NexGen HM Protease inhibitor (BPOI001) was bought from Biopioneer (India). SRL Chemicals (India) supplied the reagents, such as N, N-dimethylformamide (DMF) and N,N-diisopropylethylamine (DIPEA). Polypropylene columns were supplied by Bio-Rad. Lipofectamine 2000 was purchased from Thermo Scientific. The LDHFSR-01 kit was procured from Meril Diagnostics. VEGF antibody (sc-365578), and VEGF siRNA (sc-29520) were purchased from Santacruz. GAPDH monoclonal antibody (10-10011) and alkaline phosphatase-conjugated goat anti-mouse immunoglobulin (Ig) G (11-302) were purchased from Abgenex. 18s rRNA primer and VEGF primer (Table S3) were custom synthesized from Integrated DNA Technology (IDT), USA.

Baral B., Panigrahi B., Kar A., Tulsiyan K.D., Suryakant U., Mandal D, & Subudhi U. (2023). Peptide nanostructures-based delivery of DNA nanomaterial therapeutics for regulating gene expression. Molecular Therapy. Nucleic Acids, 33, 493-510.

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Reverse Zymography for Low-Molecular-Weight Peptides

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Reverse zymography [63 (link)] was performed using SDS-PAGE with a Tris-tricine buffer system for low molecular weight peptides [64 (link)]. An 18% polyacrylamide gel (Fisher Scientific) was co-polymerized with 0.1% (w/v) gelatin, 0.75 M Tris-HCl (pH 8.4) (Fisher Scientific) and 0.1% (w/v) SDS (MP Biomedicals) and run at a constant current of 75 V. Due to the very low molecular weight of the SFTI peptide (approx. 1.5 kDa), the 10 × 8 × 0.075 cm gel was only run ~½ of the gel length and immediately washed with 2.5% Triton X-100 (Sigma Aldrich) to remove SDS, washed with water and then digested with trypsin-containing buffer ((4.5 mg/ml trypsin in 10 mM Tris-HCl pH 7.6, 200 mM NaCl (Fisher Scientific), 10 mM CaCl₂, 0.02% (w/v) Brij-35 (Fisher Scientific)) for 3 h and then stained with Coomassie Brilliant Blue R250 (Protea Biosciences; Morgantown, WV).

Quintero D., Carrafa J., Vincent L., Kim H.J., Wohlschlegel J, & Bermudes D. (2018). Co-Expression of a Chimeric Protease Inhibitor Secreted by a Tumor-Targeted Salmonella Protects Therapeutic Proteins from Proteolytic Degradation. Journal of microbiology and biotechnology, 28(12), 2079-2094.

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Cultivation and Stress Response Assays for F. graminearum

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The wild-type strain PH-1, ectopic transformant, and Δgil1 mutant strains of F. graminearum were routinely cultured on potato dextrose agar (PDA), complete medium (CM), or 5× YEG plates at 25 °C as described [42 (link),43 (link)]. Protoplast preparation and PEG (polyethylene glycerol)-mediated transformation of F. graminearum were performed as described [5 (link),42 (link)]. To test response against various stresses, vegetative growth was assayed on CM plates with 0.7 M NaCl NaCl (Guangdong Guanghua Sci-Tech Co., Ltd., Shantou, China), 0.01% (w/v) SDS (MP Biomedicals, LLC, Solon, OH, USA), or 300 μg/mL Congo Red (SIGMA-ALDRICH Co., St. Louis, MO, USA) [13 (link),21 (link)].

Yu D., Zhang S., Li X., Xu J.R., Schultzhaus Z, & Jin Q. (2017). A Gin4-Like Protein Kinase GIL1 Involvement in Hyphal Growth, Asexual Development, and Pathogenesis in Fusarium graminearum. International Journal of Molecular Sciences, 18(2), 424.

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Tau and MAP2 Aggregation Kinetics

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Seeds (1% monomer molar equivalents of htau40 and 10% monomer equivalents of K18) were mixed with 10 μM K18 or htau40 monomer, 20 μM heparin, 0.5 to 1 mM TCEP and assembly buffer, and allowed to incubate quiescently at 37 °C for either 6 h (K18) or 21 h (htau40). Experiments probing the blockage of htau40 elongation included 10 μM 3R or 4R MAP2. Experiments probing the inhibition of K18 elongation included varying concentrations of 3R MAP2 (12.5 nM-25 μM) and 4R MAP2 (12.5 nM-15 μM) as specified in the result section. After incubation, the samples were centrifuged for 30 min at 130,000g. The pellets were separated from supernatants, volumes adjusted with sample buffer (62.5 mM TRIS pH 6.5 (Sigma), 4% SDS (J.T. Baker), 10% sucrose (MP Biomedicals), 5% 2-Mercaptoethanol, 1.5 mM Bromophenol Blue (Sigma)), and then boiled for 5 min. Equal amounts of samples were analyzed by SDS-PAGE (14–16%) and Coomassie staining. Band intensities were quantified by Image J. IC₅₀ values were computed by applying the four-parameter nonlinear regression model in GraphPad Prism 9.

Holden M.R., Krzesinski B.J., Weismiller H.A., Shady J.R, & Margittai M. (2023). MAP2 caps tau fibrils and inhibits aggregation. The Journal of Biological Chemistry, 299(7), 104891.

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Chitosan-Based Nanoparticle Synthesis

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All experiments were performed using Millipore Direct-Q 3 deionized water (18.2 MΩ·m resistivity). Chitosan (90% DD, as determined by pH titration and viscosity-average molecular weight = 120 kDa [36 (link)]), TPP (sodium salt) and NaC₁₂ were purchased from Sigma-Aldrich (St. Louis, MO, USA). SDS was obtained from MP Biomedicals (Solon, OH, USA), HCl was bought from both Sigma-Aldrich (St. Louis, MO, USA) and VWR International, LLC (West Chester, PA, USA), and NaC₁₀ was purchased from Spectrum (New Brunswick, NJ, USA). NaCl and NaOH were obtained from Fisher Scientific (Fair Lawn, NJ, USA). All materials were used as received.

Worthen A.J., Irving K.S, & Lapitsky Y. (2019). Supramolecular Strategy Effects on Chitosan Bead Stability in Acidic Media: A Comparative Study. Gels, 5(1), 11.

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