Mesna

Manufactured by Merck Group

Sourced in United States, United Kingdom

MESNA is a laboratory reagent used as a reducing agent to prevent disulfide bond formation. It is a colorless, odorless, crystalline solid that is soluble in water and alcohol. MESNA is commonly used in biochemical and biomedical research applications.

Automatically generated - may contain errors

Lab products found in correlation

Sulfo nhs ss biotin, by Thermo Fisher Scientific (4 mentions) Cyclophosphamide, by Merck Group (3 mentions) Leupeptin, by Thermo Fisher Scientific (3 mentions) Streptavidin conjugated agarose, by Merck Group (3 mentions) Iodoacetamide iaa, by Merck Group (2 mentions) B10621, by Thermo Fisher Scientific (2 mentions) 7000 mwcoslide a lyzer, by Thermo Fisher Scientific (2 mentions) Mg132, by Thermo Fisher Scientific (2 mentions) Iodoacetamide, by Merck Group (2 mentions) Ez link nhs biotin, by Thermo Fisher Scientific (2 mentions) Hispur ni nta resin, by Thermo Fisher Scientific (2 mentions) Sodium bicarbonate, by Merck Group (2 mentions) Iodoacetamide, by Bio-Rad (1 mentions) Ammonium formate, by Merck Group (1 mentions) Carvedilol, by Merck Group (1 mentions) Morbital, by Biowet (1 mentions) Afatinib, by Selleck Chemicals (1 mentions) Penicillamine, by Merck Group (1 mentions) Minocycline, by Merck Group (1 mentions) Y1068, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Sodium 2-mercaptoethane sulfonate (mesna ; (2 citations)

29 protocols using mesna

Preparation and Characterization of ISG15-USP18 Conjugates

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hISG15(79-C)-propargylamide(PA) (hISG15-PA) and mISG15(77-C)-PA (mISG15-PA) probes were prepared with Intein-MESNa (sodium 2-mercaptoethanesulfonate) methodology similar to the previous reports.⁵² (link) Briefly, ISG15-Intein-CBS proteins were expressed in BL21(DE3) and purified by Chitin-Binding beads (NEB, S6651) affinity purification followed by eluting with SEC buffer (25 mM HEPES pH 8.0, 200 mM NaCl, 1 mM TCEP) containing 50mM MESNa (Sigma,1392807). ISG15-MESNa conjugates were further purified by size exclusion chromatography with the SEC buffer followed by converting it to ISG15-PA by incubating with 5 times molar PA (Sigma, P50900) at RT. Excess PA was removed by desalting using PD10 desalting column (Cytiva, 17085101).
50 nM hUSP18 WT and 5000 nM hISG15-PA or 1 μM mUSP18 WT and 10 μM mISG15-PA were incubated at 37°C for 30 min to monitor the conjugation of hUSP18-hISG15 or mUSP18-mISG15, respectively. hUSP18-hISG15 conjugation was detected by western blot with the following primary antibody: rabbit anti-USP18 (Cell Signaling, 4813S). mUSP18-mISG15 conjugation was visualized by trihalo compound-based, stain-free SDS-PAGE (BioRad, 4568086).

Jové V., Wheeler H., Lee C.W., Healy D.R., Levine K., Ralph E.C., Yamaguchi M., Jiang Z.K., Cabral E., Xu Y., Stock J., Yang B., Giddabasappa A., Loria P., Casimiro-Garcia A., Kessler B.M., Pinto-Fernández A., Frattini V., Wes P.D, & Wang F. (2024). Type I interferon regulation by USP18 is a key vulnerability in cancer. iScience, 27(4), 109593.

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Cell Surface Biotinylation Assay

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Cells were seeded overnight at the above described densities. The following day media was changed and cells were incubated at 37 °C for 24 h. Subsequently, cells were washed 1× with PBS and 125 μL of 0.2 mg/mL cell permeable NHS-SS-Biotin (Pierce) was added to each well and incubated for 1 h at 4 °C with gentle agitation. Cells were then washed 1× with Reducing Buffer (50 mM Tris, 150 mM NaCl, pH 8.6) and cells were either stripped with the addition of 20 mM sodium 2-mercaptoethanesulfonate (MESNA, Sigma) or non-stripped with Reducing buffer and incubated for 1 h at 4 °C with gentle agitation. The stripping reaction was quenched by addition of 40 mM Iodoacetamide (IAA, Sigma) for 10 min at 4 °C with gentle agitation. Cells were washed 1× with PBS and lysed. Lysates were measured for total biotin signal (not stripped with MESNA) and internal biotin signal (surface stripped with MESNA) with an adapted Luminex protocol, eliminating the detection antibody step.

Claas A.M., Atta L., Gordonov S., Meyer A.S, & Lauffenburger D.A. (2018). Systems Modeling Identifies Divergent Receptor Tyrosine Kinase Reprogramming to MAPK Pathway Inhibition. Cellular and Molecular Bioengineering, 11(6), 451-469.

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Evaluating Boswellic Acids and Cyclophosphamide

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All the chemicals
used were of analytical grade. Both cyclophosphamide and mesna (Sigma-Aldrich)
were obtained from the Oncology Department, Doctors Hospital Lahore,
Lahore, and CYP was reconstituted with 0.9% normal saline to make
an injectable solution. Boswellic acids (97%) was obtained from Yuantai
Biological Technology (China).

Fatima M., Anjum I., Abdullah A., Abid S.Z, & Malik M.N. (2022). Boswellic Acids, Pentacyclic Triterpenes, Attenuate Oxidative Stress, and Bladder Tissue Damage in Cyclophosphamide-Induced Cystitis. ACS Omega, 7(16), 13697-13703.

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Surface Protein Biotinylation and Degradation

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Surface biotinylation was performed as described elsewhere (Gottardi et al., 1995 (link)). In brief, cells were rinsed in PBS without calcium or magnesium (PBS⁻) and labeled at 4°C for 30 min with 0.5 mg/ml sulfo-NHS-SS biotin (Pierce) in PBS⁻. The reaction was quenched with 20 mM glycine in PBS⁻. To assess the degradation kinetics of biotinylatable (surface) proteins, cells were placed at 37°C to induce endocytosis (and subsequent degradation) for various time points, washed with PBS⁻, and lysed with 1% Triton buffer described earlier. Biotinylated proteins were then enriched by affinity purification with NeutrAvidin beads (Pierce). To assess endocytosis rates, labeled cells were returned to 37°C for various time points but were then placed back on ice. Remaining biotin on the cell surface was stripped with MesNA (Sigma-Aldrich) in 50 mM Tris, pH 8.6, 100 mM NaCl, and 2.5 mM CaCl₂ followed by an alkylation step with 5 mg/ml iodoacetamide (Bio-Rad) in PBS⁻. Cells were washed with PBS⁻ before lysis and enrichment as described. One plate for each condition was not placed at 37°C and served as a control for total labeling efficiency, and an additional plate was not placed at 37°C but was subjected to the stripping and alkylation steps as a control for stripping efficiency.

McEwen A.E., Maher M.T., Mo R, & Gottardi C.J. (2014). E-cadherin phosphorylation occurs during its biosynthesis to promote its cell surface stability and adhesion. Molecular Biology of the Cell, 25(16), 2365-2374.

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Site-specific Protein Labeling with Bifunctional Rhodamine

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Attachment of bifunctional rhodamine (Bis-((N-Iodoacetyl)-Piperazinyl)-
Sulfonerhodamine; Invitrogen B10621) with two iodoacetamide groups to E146C and
L153C in the regulatory module and N-terminal biotinylation were performed
following a previously described protocol with some modifications^{8 (link)}. The protein sample in the gel
filtration buffer at an ~50 μM concentration was treated with 50
μM reducing agent TCEP (Sigma T2556) at room temperature for one hour,
after which 30 μM bifunctional rhodamine was added four times at
12-minute intervals. 54 minutes after the fourth addition, the labeling reaction
was terminated with 3 mM MESNA (Sigma-Aldrich, M1511). As high concentration of
K⁺ inhibits the biotin ligase (BiRA; Avidity), the above protein
sample was diluted 2.5 times with a solution containing 100 mM NaCl, and 2 mM
EDTA titrated to pH 8.0, lowering the K⁺ concentration to 100 mM.
After adding a manufacturer-supplied reaction mix and BiRA (40 μg per 10
nmoles of sample protein), the biotin ligation reaction proceeded for 1 hour at
30 °C. To remove free biotin and bifunctional rhodamine and to raise KCl
back to 250 mM KCl, the sample was dialyzed overnight in a 7000 MWCO
Slide-A-lyzer (ThermoFisher 66373) against a solution containing 250 mM KCl and
10 mM Tris titrated to pH 8.0, in a 1:1000 volume ratio.

Lewis J.H, & Lu Z. (2019). Resolution of angstrom-scale protein-conformational changes by analyzing fluorescence anisotropy. Nature structural & molecular biology, 26(9), 802-807.

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Site-specific Protein Labeling with Bifunctional Rhodamine

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Lewis J.H, & Lu Z. (2019). Resolution of angstrom-scale protein-conformational changes by analyzing fluorescence anisotropy. Nature structural & molecular biology, 26(9), 802-807.

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Biotinylation and Internalization Assay

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Cells pretreated with 10μM MG132 and 50μM Leupeptin (FisherScientific) were labeled at 4°C with Sulfo-NHS-S-S-biotin (ThermoFisher Scientific) in PBS, followed by incubation at 37°C for internalization of surface molecules. Residual surface biotinylation was removed by incubation with 200mM MESNA (Sigma) for 30 minutes. Cells were homogenized in PTY buffer (62 (link)), and biotinylated molecules were pulled down with streptavidin-conjugated agarose (Millipore) and used for western blotting.

Loskutov Y.V., Kozyulina P.Y., Kozyreva V.K., Ice R.J., Jones B.C., Roston T.J., Smolkin M.B., Ivanov A.V., Wysolmerski R.B, & Pugacheva E.N. (2014). NEDD9/Arf6-Dependent Endocytic Trafficking of Matrix Metalloproteinase 14: A Novel Mechanism for Blocking Mesenchymal Cell Invasion and Metastasis of Breast Cancer. Oncogene, 34(28), 3662-3675.

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MESNA and 3-MPS Quantification Protocol

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All reagents and solvents were at least HPLC grade, unless otherwise stated. Ammonium formate was purchased from Sigma-Aldrich (St. Louis, MO, USA). Methanol and acetone (LC–MS grade) were obtained from Fisher Scientific (Hanover Park, IL, USA). MESNA (>98% purity) and the internal standard, sodium 3-mercapto-1-propane sulfonate (3-MPS, >90% purity), were purchased from Sigma-Aldrich (St. Louis, MO, USA). Water used for this study was purified by reverse osmosis and filtered through a Lab Pro polishing unit from Labconco (Kansas City, KS, USA). Stock solutions of MESNA and 3-MPS (10 mM) were prepared in water and stored at 4 °C. The stock solutions of MESNA and 3-MPS were freshly diluted to desired working solutions in water or plasma immediately prior to each experiment.

Donkor A.B., White C.W., Nick H.J, & Logue B.A. (2021). Analysis of sodium 2-mercaptoethane sulfonate in rat plasma using high performance liquid chromatography tandem-mass spectrometry. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 1189, 123088.

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Pharmacological Cocktail for Experimental Treatment

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Cyclophosphamide subst. (Sigma, Steinheim, Germany), mesna subst. (Sigma, Steinheim, Germany), carvedilol subst. (Sigma, Steinheim, Germany), 0.9% NaCl solution, ampules 10 mL (Polpharma S.A. Starogard Gdański, Poland), pentobarbital sodium 133.3 mg/mL + pentobarbital 26.7 mg/mL, bottles 100 mL (Morbital^®, Biowet, Puławy, Poland).

Merwid-Ląd A., Ziółkowski P., Szandruk-Bender M., Matuszewska A., Szeląg A, & Trocha M. (2021). Effect of a Low Dose of Carvedilol on Cyclophosphamide-Induced Urinary Toxicity in Rats—A Comparison with Mesna. Pharmaceuticals, 14(12), 1237.

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EGFR Pathway Modulation Assay

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Bucillamine was purchased from FUJIFILM Wako Chemicals USA Corporation (Richmond, VA, USA) or Santa Cruz Biotechnology (Dallas, TX, USA); penicillamine, MESNA, and minocycline from Sigma; afatinib and afatinib dimaleate from Selleck (Houston, TX, USA. Antibodies against EGFR and pEGFR (Y1068) were purchased from Cell Signaling Technology (Beverly, MA, USA). Horseradish peroxidase (HRP)-conjugated anti-rabbit immunoglobulin G (IgG) and anti-mouse IgG antibodies were purchased from GE Healthcare (Chicago, IL, USA). The A431 cell line was obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 2 mM L-glutamine and 10% fetal bovine serum in a humidified 5% CO₂ atmosphere at 37 °C.

Nishiya N., Murai M., Hosoda A., Yonezawa H, & Omori N. (2019). Bucillamine Prevents Afatinib-Mediated Inhibition of Epidermal Growth Factor Receptor Signaling. Pharmaceuticals, 12(4), 165.

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