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Guanidine hydrochloride guhcl

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Guanidine hydrochloride (GuHCl) is a chemical compound that is commonly used in laboratory settings. It is a strong chaotropic agent, meaning it can disrupt the structure of proteins and nucleic acids. GuHCl is often used in the process of protein denaturation and purification.

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Lab products found in correlation

D glucose, by Merck Group (2 mentions) 96 well plate, by Greiner (2 mentions) Renilla luciferase assay system, by Promega (2 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions) Cycloheximide (chx), by Merck Group (2 mentions) Renilla luciferase assay lysis buffer, by Promega (2 mentions) Dipyridamole, by Merck Group (2 mentions) Milliq direct q uv, by Merck Group (1 mentions) Ammonia solution, by Merck Group (1 mentions) D fructose, by Merck Group (1 mentions) D sucrose, by Merck Group (1 mentions) D galactose, by Merck Group (1 mentions) Lc ms lichropur grade, by Merck Group (1 mentions) Amicon ultra 0.5 centrifugal filter unit 3 kda, by Merck Group (1 mentions) D lactose monohydrate, by Merck Group (1 mentions) Millex lcr filters, by Merck Group (1 mentions) Tris 2 carboxyethyl phosphine tcep, by Merck Group (1 mentions) Acetonitrile, by Merck Group (1 mentions) Triisopropylsilane, by Merck Group (1 mentions) N dimethylformamide, by Merck Group (1 mentions)

Close spelling variants of this product

Guanidine hydrochloride (194 citations) GuHCl (21 citations) Hydrochlorides (2 citations)

15 protocols using guanidine hydrochloride guhcl

1

Carbohydrate Profiling: Standards and Preparation

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Standards of mono- and disaccharides: d-fructose, d-glucose, d-galactose, d-(+)-sucrose, d-lactose monohydrate, and ammonia solution (25%, LC-MS LiChropur™ grade) were obtained from Sigma-Aldrich (Darmstadt, Germany). Glucose-13C6 (Glu-13C6, U-13C6, 99%, chemical purity 98%) and lactose monohydrate (Lac-13C6, UL-13C6glc, 98%+) were procured from Cambridge Isotope Laboratories Inc. (Tewksbury, MA, USA). Ultrapure water (18.2 MΩ.cm) was prepared with MilliQ® Direct-Q® UV (Merck KGaA, Darmstadt, Germany). Acetonitrile (MeCN; LiChrosolv, HPLC gradient grade), and guanidine hydrochloride (GuHCl; ≥99%) were acquired from Sigma-Aldrich (Darmstadt, Germany). Biotage Isolute® PLD+, C18 and NH2 were procured from Biotage Sweden AB (Uppsala, Sweden). Amicon Ultra-0.5 centrifugal filter unit (3 kDa) and Millex-LCR filters (Pore size 0.2 µm, Filter Dimension 13 mm) were obtained from Merck KGaA (Darmstadt, Germany) and Microsep Advance Centrifugal Devices with Omega Membrane 1K filter unit was purchased from Pall Corporation (Port Washington, NY, USA).
Pismennõi D., Kiritsenko V., Marhivka J., Kütt M.L, & Vilu R. (2021). Development and Optimisation of HILIC-LC-MS Method for Determination of Carbohydrates in Fermentation Samples. Molecules, 26(12), 3669.
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2

IgG Antibody Avidity Assay for HPV

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To approximate the binding strength of IgG antibodies to HPV, the avidity of antibodies was determined as previously described, using a modified ELISA-based method with the chaotropic agent guanidine hydrochloride (GuHCl; Sigma, St. Louis, MO) [20 (link),21 (link)]. The avidity index is the concentration of GuHCl, expressed in Molar (M), that reduce the optical density by 50% compared to sample wells without GuHCl treatment. Flat-bottom microtiter plates were coated with HPV antigen, and blocked, as previously described above in the HPV ELISA method. Serum samples were diluted based on previous evaluation in the HPV-16 or HPV-18 ELISA to yield an absorbance reading of 1.0 ± 0.5. Samples were excluded if their reading was below 1.0 ± 0.5, or the sample could not be diluted down to 1.0 ± 0.5 with a 1:100 dilution. The diluted serum was incubated for 1 h at room temperature. Buffer alone, or 0.5 to 3.5 M GuHCl was added for 15 min at room temperature. After washing the plate, the secondary goat anti-human IgG was added for one hour at room temperature, and the plate was developed with TMB and the absorbance was read as previously described for the HPV ELISA.
Miller C.N., Kemp T.J., Abrahamsen M., Isaacs-Soriano K., Dunham K., Sirak B., Pan Y., Lazcano-Ponce E., Salmeron J., Pinto L.A, & Giuliano A.R. (2021). Increases in HPV-16/18 antibody avidity and HPV-specific memory B-cell response in mid-adult aged men post-dose three of the quadrivalent HPV vaccine. Vaccine, 39(37), 5295-5301.
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3

Synthesis of Lipid-Peptide Conjugates

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Soybean lecithin, DSPE-PEG2000 and DSPE-PEG2000-Maleimide was purchased from Xi’an Ruixi Biological Technology Co., Ltd, oridonin from MCE®, and DCM from Sigma-Aldrich®. TLR2 pep-Cys was chemically synthesized on solid phase. P-methyl-BHA (MBHA) resin was purchased from Applied Biosystems®, Boc protected amino acids were purchased from Peptides Institute®. Other reagents for peptide synthesis and cleavage, including GSH, dichloromethane (DCM), N, N-dimethylformamide (DMF), acetonitrile, Tris-(2-carboxyethyl) phosphine (TCEP), triisopropylsilane (TIPS), N, N-diisopropylethylamine (DIEA), p-cresol and guanidine hydrochloride (GuHCl) were purchased from Sigma-Aldrich®. Trifluoroacetic acid (TFA) was purchased from Halocarbon®.
Liu Y., Wang X., Feng H., Li X., Yang R., Zhang M., Du Y., Liu R., Luo M., Li Z., Liu B., Wang J., Wang W., An F., Niu F, & He P. (2024). Glutathione-depleting Liposome Adjuvant for Augmenting the Efficacy of a Glutathione Covalent Inhibitor Oridonin for Acute Myeloid Leukemia Therapy. Journal of Nanobiotechnology, 22, 299.
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4

Fluoxetine and Guanidine Hydrochloride Protocol

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HEp-2 cells (BioWhittaker, Walkersville, MD, USA) were grown in minimum essential medium (MEM) supplemented with 10% of fetal calf serum (FCS), 1% of L-glutamine, 1% of nonessential amino acids, and 1% of penicillin and streptomycin. The human ductal cell line Panc-1 (ATCC) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% of FCS, 1% of L-glutamine, and 1% of penicillin and streptomycin.
Fluoxetine chlorhydrate (Lilly France, Fegersheim, France) was dissolved in dimethyl sulfoxide (DMSO) at a final concentration of 5.48 uM and was used in all experiments, as previously reported [12 (link)]. Guanidine hydrochloride (GuHCl) was purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France) and was used at a final concentration of 2 mM.
Alidjinou E.K., Bertin A., Sane F., Caloone D., Engelmann I, & Hober D. (2019). Emergence of Fluoxetine-Resistant Variants during Treatment of Human Pancreatic Cell Cultures Persistently Infected with Coxsackievirus B4. Viruses, 11(6), 486.
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5

High-Fat Diet Alters Gastrointestinal Proteome

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Male mice were fed either a high-fat diet (HFD) (60 % kcal fat, Research Diets (New Brunswick, NJ, USA), D12492) or a standard laboratory chow diet for 13 weeks. At the end of the 13 weeks, fasting blood glucose levels were measured following a 6 h fast. Mice were culled and the GI tracts dissected out. Pieces (∼30 mg) of mouse GI mucosa were flushed with PBS and homogenised in 250 μL 6 M guanidine hydrochloride (GuHCl) (Sigma-Aldrich) with Lyzing MatrixD (MP biomedicals) beads using a FastPrep-24 homogeniser. 4 cycles of 40 s shaking at 6 m/s was used to homogenise tissue. Proteins in samples were precipitated by addition of 80 % ACN and centrifuged (12 000 g, 5 min, 4⁰C). The aqueous phase was removed and dried overnight in a centrifugal evaporator.
Galvin S.G., Larraufie P., Kay R.G., Pitt H., Bernard E., McGavigan A.K., Brant H., Hood J., Sheldrake L., Conder S., Atherton-Kemp D., Lu V.B., O’Flaherty E.A., Roberts G.P., Ämmälä C., Jermutus L., Baker D., Gribble F.M, & Reimann F. (2021). Peptidomics of enteroendocrine cells and characterisation of potential effects of a novel preprogastrin derived-peptide on glucose tolerance in lean mice. Peptides, 140, 170532.
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6

Measuring Viral Entry Inhibition

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Cells were seeded on 96-well plates (Greiner Bio-One) in triplicates and incubated for 16 to 24 hours. Cells were first pre-chilled on ice for 30 minutes. Next, CV-B3-RLuc (MOI=10) were pre-bound to cells on ice for 60 minutes in the presence of either dimethyl sulfoxide (DMSO, Thermo Fisher) at a final concentration of 0.1%, guanidine hydrochloride (GuHCl, Sigma-Aldrich) at a final concentration of 2 mM or cycloheximide (CHX, Sigma-Aldrich), at a final concentration of 125 μM. The cells were then incubated at 37°C in incubator for 10 minutes to permit viral entry. Next, the cells were washed 3 times with PBS and cultured in complete culture media supplemented with DMSO, GHCl, or CHX at the above-mentioned final concentration. At indicated time points, total cell lysates were harvested in Renilla Luciferase Assay lysis buffer and luciferase expression was measured using Renilla Luciferase Assay system (Promega E2820). Luciferase activity was measured by addition of substrate and luciferase readings were taken immediately using Glomax 20/20 luminometer using a 5 second integration time.
Diep J., Ooi Y.S., Wilkinson A.W., Peters C.E., Foy E., Johnson J.R., Zengel J., Ding S., Weng K.F., Laufman O., Jang G., Xu J., Young T., Verschueren E., Kobluk K.J., Elias J.E., Sarnow P., Greenberg H.B., Hüttenhain R., Nagamine C.M., Andino R., Krogan N.J., Gozani O, & Carette J.E. (2019). Enterovirus Pathogenesis Requires the Host Methyltransferase SETD3. Nature microbiology, 4(12), 2523-2537.
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7

Protein Biophysical Characterization

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Bovine carbonic anhydrase II (BCAII), hen egg white lysozyme, Ribonuclease A, Micrococcus lysodeikticus, cystine hydrochloride, fluorescein isothiocyanate (FITC), Guanidine hydrochloride (GuHCl), dithiothreitol (DTT), GTP, ATP and antibiotics that specifically bind to domainV of 23S rRNA (blasticidin and chloramphenicol) were purchased from Sigma. Nitrocellulose filter was purchased from Millipore, p-nitrophenylacetate (p-NPA) from SRL biochemical and reagents for molecular biology like T7 RNA Polymerase and RNase free DNase I were purchased from Fermentas, Ni+2-NTA was purchased from Qiagen. All other chemicals were local products of analytical grade. All data analysis was performed using OriginPro 8 software.
Pathak B.K., Mondal S., Ghosh A.N, & Barat C. (2014). The Ribosome Can Prevent Aggregation of Partially Folded Protein Intermediates: Studies Using the Escherichia coli Ribosome. PLoS ONE, 9(5), e96425.
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8

Optimized Protease Assay Buffer Preparation

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Use an optimized assay buffer for the target protease, if known. Detergents should be avoided whenever possible due downstream ionizing effects in the mass spectrometer that will decrease the signal of the peptides. If an optimized buffer is not known, some common assay buffers that may be used include:

20 mM Citrate Phosphate, pH 3.5, 100 mM NaCl

20 mM Citrate Phosphate, pH 5.5, 100 mM NaCl

20 mM Tris–HCl, pH 7.5, 100 mM NaCl

8 mM Na2HPO4, 2 mM KH2PO4, 137 mM NaCl, 2.7 mM KCl (phosphate buffered saline)

8.0 mM Na2HPO4, 1.47 mM KH2PO4, 138 mM NaCl, 2.7 mM KCl, 0.5 mM MgCl2, 0.9 mM CaCl2 (Dulbecco’s phosphate buffered saline)

For sample quenching, desalting and LC-MS/MS, the following solvents are used

8 M Guanidine hydrochloride (GuHCl) (Sigma)

8 M Urea (Sigma)

Acetonitrile (ACN), LC-MS grade (Thermo)

Formic acid (FA), LC-MS grade (Thermo)

Methanol, LC-MS grade (Thermo)

Trifluoracetic acid (TFA), LC-MS grade (Thermo)

Water, LC-MS grade (Thermo)

Solvent A: 0.1% FA/H2O or 0.1% TFA/H2O

Solvent B: 0.1% FA in 50% ACN/H2O or 0.1% TFA in 50% ACN/H2O

Solvent C: 0.1% FA in 70% ACN/H2O or 0.1% TFA in 70% ACN/H2O

Rohweder P.J., Jiang Z., Hurysz B.M., O’Donoghue A.J, & Craik C.S. (2022). Multiplex substrate profiling by mass spectrometry for proteases. Methods in enzymology, 682, 375-411.
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9

IgG Antibody Avidity Assay for HPV

Cited 2 times
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To approximate the binding strength of IgG antibodies to HPV, the avidity of antibodies was determined as previously described, using a modified ELISA-based method with the chaotropic agent guanidine hydrochloride (GuHCl; Sigma, St. Louis, MO) [20 (link),21 (link)]. The avidity index is the concentration of GuHCl, expressed in Molar (M), that reduce the optical density by 50% compared to sample wells without GuHCl treatment. Flat-bottom microtiter plates were coated with HPV antigen, and blocked, as previously described above in the HPV ELISA method. Serum samples were diluted based on previous evaluation in the HPV-16 or HPV-18 ELISA to yield an absorbance reading of 1.0 ± 0.5. Samples were excluded if their reading was below 1.0 ± 0.5, or the sample could not be diluted down to 1.0 ± 0.5 with a 1:100 dilution. The diluted serum was incubated for 1 h at room temperature. Buffer alone, or 0.5 to 3.5 M GuHCl was added for 15 min at room temperature. After washing the plate, the secondary goat anti-human IgG was added for one hour at room temperature, and the plate was developed with TMB and the absorbance was read as previously described for the HPV ELISA.
Miller C.N., Kemp T.J., Abrahamsen M., Isaacs-Soriano K., Dunham K., Sirak B., Pan Y., Lazcano-Ponce E., Salmeron J., Pinto L.A, & Giuliano A.R. (2021). Increases in HPV-16/18 antibody avidity and HPV-specific memory B-cell response in mid-adult aged men post-dose three of the quadrivalent HPV vaccine. Vaccine, 39(37), 5295-5301.
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10

SARS-CoV-2, HCoV-OC43, and H1N1 RNA Detection

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Genomic SARS-CoV-2 RNA (2019-nCoV/USA-WA1/2020, ATCC VR-1986D), HCoV-OC43 (ATCC VR-1558D) and H1N1 (ATCC VR-1736D) were purchased from LGC standards, UK. WarmStart® DNA and RNA polymerase (MS1800S) was purchased from New England Biolabs. Primers were purchased from Integrated DNA technologies (IDT). Oligo (dT)-coated magnetic beads, SYBR Safe and nuclease-free water were supplied by Thermofisher Scientific, UK. Mineral oil and guanidine hydrochloride (GuHCl) were purchased from Sigma-Aldrich.
Rodriguez-Mateos P., Ngamsom B., Walter C., Dyer C.E., Gitaka J., Iles A, & Pamme N. (2021). A lab-on-a-chip platform for integrated extraction and detection of SARS-CoV-2 RNA in resource-limited settings. Analytica Chimica Acta, 1177, 338758.
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