2n sulfuric acid

Manufactured by Merck Group

2N sulfuric acid is a commonly used laboratory reagent. It is an aqueous solution of sulfuric acid with a normality of 2, meaning it has a concentration of 2 equivalents of hydrogen ions per liter of solution. This solution is typically used for various analytical, purification, and reaction processes in laboratory settings.

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Lab products found in correlation

Octectred system, by Molecular Devices (2 mentions) Horseradish peroxidase conjugated goat anti human igg fc, by Southern Biotech (2 mentions) Tmb elisa chromogenic substrate, by Thermo Fisher Scientific (2 mentions) Maxisorp, by Thermo Fisher Scientific (2 mentions) Blotto, by Thermo Fisher Scientific (2 mentions) Tmb substrate solution, by BioLegend (1 mentions) 96 well plate, by Corning (1 mentions) Spectramax 190 microplate reader, by Molecular Devices (1 mentions) Multiskan skyhigh photometer, by Thermo Fisher Scientific (1 mentions) Tetramethylbenzidine solution, by Merck Group (1 mentions) Sodium bicarbonate, by Merck Group (1 mentions) Peroxidase conjugated secondary antibody, by Vector Laboratories (1 mentions) Sodium carbonate, by Merck Group (1 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by BD (1 mentions) Uquant microplate reader, by Agilent Technologies (1 mentions) Pbs tween 20, by Merck Group (1 mentions) Bsa pbs, by Merck Group (1 mentions) Tmb solution, by Thermo Fisher Scientific (1 mentions) Victor x3 plate reader, by PerkinElmer (1 mentions)

Spelling variants of this product

Sulfuric acid (1309 citations)

7 protocols using 2n sulfuric acid

Quantification of OGT and O-GlcNAc

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For the detection of OGT or O-GlcNAc, 96-well plates (Corning) were coated with lysates from pre-B cells transfected by siOGT or siCONTROL, serially diluted in PBS, at 4 °C overnight. The plates were washed thrice with PBS-T (0.05% Tween 20 in 1× PBS) and incubated in blocking buffer (0.1% Tween 20 and 1% BSA in PBS) at RT for 2 h, followed by PBS-T-washes three times. The plates were washed five times with PBS-T and treated with a primary antibody, anti-OGT, or anti-O-GlcNAc (CTD110.6), diluted 1:1000 in an assay buffer (0.05% Tween 20 and 0.05% BSA in PBS) at RT for 2 h, followed by PBS-T washes five times. Then, detecting antibody (HRP-conjugated anti-mouse IgG or IgM) was diluted 1:5000 in an assay buffer at RT for 1 h. For the detection of c-Myc, 96-well plates were initially coated with 1 μg/mL of capture antibody, anti-c-Myc, at 4 °C overnight, and then cell lysates were applied to the plate after blocking. HRP-conjugated anti-Myc antibody was used as a detecting antibody. After five washes with PBS-T post treatment of each detecting antibody, TMB Substrate Solution (BioLegend, San Diego, CA, USA, 77247 & 77248) was added to the plates. After an appropriate time of incubation, the reaction was stopped by adding 2 N sulfuric acid (MilliporeSigma). The plates were finally read at 450 nm using a SpectraMAX 190 Microplate Reader (Molecular Devices).

Lee D.H., Kwon N.E., Lee W.J., Lee M.S., Kim D.J., Kim J.H, & Park S.K. (2020). Increased O-GlcNAcylation of c-Myc Promotes Pre-B Cell Proliferation. Cells, 9(1), 158.

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Monoclonal Antibody Binding to Mycobacterium Antigen

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MAb binding to AM assay was performed by BLI using the OctectRed system (ForteBio, Pall) (33 (link)). Briefly, purified capsular AM was biotinylated and immobilized onto streptavidin-coated sensors. The sensor was then dipped into wells containing 1 mAb at serial dilutions to create binding curves and calculate the K_D. The binding of AM mAbs to purified capsular AM and intact bacterial surface was further quantified by ELISA (33 (link)). Briefly, purified AM was coated on 96-well microtiter plates (Maxisorp, Thermo Fisher Scientific) at 10 μg/mL in 50 μL. For whole-cell ELISA, M. tuberculosis strains grown in media without detergent were coated on 96-well plates overnight and blocked with BLOTTO (Thermo Fisher Scientific) (33 (link)). Serially diluted mAbs were then added to the antigen- or bacteria-coated wells, and the bound mAb was probed with horseradish peroxidase–conjugated goat anti-human IgG Fc (2048-05, SouthernBiotech), followed by TMB-ELISA chromogenic substrate (Thermo Fisher Scientific). The reaction was stopped by adding an equal volume of 2N sulfuric acid (MilliporeSigma), and the optical densities (ODs) were measured at 450 nm subtracted by 540 nm. A human IgG1 mAb against a non–M. tuberculosis antigen was included as a negative control.

Liu Y., Chen T., Zhu Y., Furey A., Lowary T.L., Chan J., Bournazos S., Ravetch J.V, & Achkar J.M. (2023). Features and protective efficacy of human mAbs targeting Mycobacterium tuberculosis arabinomannan. JCI Insight, 8(20), e167960.

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Monoclonal Antibody Binding to Mycobacterium Antigen

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ZIKV and DENV Antibody Detection ELISA

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Serum samples from mice born to ZIKV- or DENV-immune mothers were tested for ZIKV- and DENV-binding antibodies using a direct ELISA. To detect DENV antibodies, sucrose purified DENV2 S221 virions were used at a concentration of 1 × 10⁶ FFU/well as the coating antigen and UV-inactivated. DENV2 was diluted in 50 μL coating buffer (0.1M NaHCO₃ in PBS) per well and incubated at 4°C overnight. Wells were washed 3 times with ELISA washing buffer (0.05% Tween20 in PBS) and then blocked with 5% casein in PBS for an hour at room temperature. Serum was first diluted 1:10 in 10% FBS/PBS and then 1:3 for subsequent dilutions and was incubated in wells for 1.5 hours at room temperature. Wells were washed 3 times with ELISA washing buffer. Wells were then incubated with peroxidase conjugated Affini-Pure Goat anti-mouse IgG Fcγ (Jackson ImmunoResearch, cat. #115-035-008) diluted 1:5000 in 1% BSA/PBS at room temperature. Wells were washed 3 times with ELISA washing buffer. 100 μL of TMB substrate solution was added until blue color change, and reaction was stopped with 50 μL of 2N sulfuric acid (Sigma, cat. #339741). To detect ZIKV Abs, ZIKV E protein (Suriname strain, Native Antigen Company, #ZIKVSU-ENV) was adsorbed to 96-well plates at a concentration of 1 μg/mL in coating buffer (0.1M NaHCO₃ in PBS) overnight at 4 °C and the remaining steps were the same as the DENV ELISA.

Fowler A.M., Tang W.W., Young M.P., Mamidi A., Viramontes K.M., McCauley M.D., Carlin A.F., Schooley R., Swanstrom J., Baric R.S., Govero J., Diamond M.S, & Shresta S. (2018). Maternally Acquired Zika Antibodies Enhance Dengue Disease Severity in Mice. Cell host & microbe, 24(5), 743-750.e5.

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Indirect ELISA for ACTH and Cortisol

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The amounts of ACTH and cortisol were measured in the serum samples described in 2.5.1 via an indirect-ELISA. Briefly, 96-well microplates (LPS solution, Daejeon, Republic of Korea) were coated with a buffer (0.3% sodium carbonate (Sigma-Aldrich), 0.6% sodium bicarbonate (Sigma-Aldrich), and 91.9% distilled water) and left overnight at 4 °C. The coated microplates were washed with phosphate-buffered saline containing 0.1% Tween-20 (TPBS; LPS solution) and blocked with 5% skim milk (LPS solution) for 6 h at room temperature. After being washed with TPBS, 100 μg of serum was added into each well and then incubated overnight at 4 °C. After washing with TPBS, the microplates were incubated with anti-ACTH or anti-cortisol antibodies overnight at 4 °C (Table S1). Following washing with TBPS, the primary antibodies were incubated with a peroxidase-conjugated secondary antibody (Vector Laboratories, Burlingame, CA, USA) at room temperature for 4 h. After another round of washing, a tetramethylbenzidine solution (Sigma-Aldrich) was added and allowed to incubate in the dark at room temperature for 15–20 min to enable color development. To terminate the reaction, an equal volume of 2 N sulfuric acid (Sigma-Aldrich) was added. The optical density at 450 nm was measured using a microplate reader (Multiskan SkyHigh Photometer; Thermo Fisher Scientific).

Batsukh S., Oh S., Rheu K., Lee B.J., Choi C.H., Son K.H, & Byun K. (2023). Rice Germ Attenuates Chronic Unpredictable Mild Stress-Induced Muscle Atrophy. Nutrients, 15(12), 2719.

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ELISA Quantification of IgM and IgG

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Total IgM and IgG levels in culture supernatants were quantified by ELISA (Bethyl Laboratories, Montgomery, TX), developed with TMB (BD Biosciences, San Jose, CA), stopped using 2N sulfuric acid (Sigma-Aldrich, St. Luis, MO), and read using the uQuant Microplate Reader (Bio-Tek, Winooski, VT).

Simchoni N, & Cunningham-Rundles C. (2015). TLR7 and TLR9 responsive human B cells share phenotypic and genetic characteristics. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3035-3044.

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Plasma Antibody Quantification by ELISA

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For plasma antibody levels, whole blood was obtained from mice by means of cardiac puncture during euthanasia and diluted 1:5 in 1% heparin-PBS (Thermo Fisher Scientific). Plasma was separated by means of centrifugation and then stored at 2808C until ELISA analysis. Ninety-six-well ELISA plates (Costar, Corning Life Sciences) were coated with antigen (20 mg/mL for bacterial lysate or OVA or 2-5 mg/mL for rP1 C-adhesin C-terminal peptide) overnight in antigen coating buffer (100 mmol/L bicarbonate/carbonate solution, pH 9.6) at 48C. In brief, antigen solution was decanted, and plates were blocked with 1% BSA-PBS (Sigma-Aldrich) solution for 1 hour at room temperature. Plasma was diluted 1:50 in 1% BSA-PBS and incubated for 2 hours at 378C. Plates were washed 3 times with 0.05% Tween-20-PBS (Sigma-Aldrich) before secondary antibody addition. Secondary antibody solution was incubated at room temperature for 1 hour. Plates were washed an additional 3 times before TMB solution addition (Thermo Fisher Scientific), according to the manufacturer's specifications. HRP substrate reactions were halted with 2N sulfuric acid (Sigma-Aldrich), and absorbance was measured at 405 nm by using a Victor X3 Plate Reader (PerkinElmer, Waltham, Mass).

Totten A.H., Xiao L., Luo D., Briles D., Hale J.Y., Crabb D.M., Schoeb T.R., Alishlash A.S., Waites K.B, & Atkinson T.P. (2019). Allergic airway sensitization impairs antibacterial IgG antibody responses during bacterial respiratory tract infections. The Journal of allergy and clinical immunology, 143(3).

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