Anti igg2c

Manufactured by Southern Biotech

Sourced in United States

Anti-IgG2c is a laboratory reagent that specifically binds to the IgG2c subclass of immunoglobulins. It can be used to detect, measure, or isolate IgG2c molecules in various experimental or analytical applications.

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Lab products found in correlation

Anti igg2b, by Southern Biotech (3 mentions) Streptavidin alkaline phosphatase, by Vector Laboratories (2 mentions) Poly l lysine (pll), by Merck Group (2 mentions) Salmon sperm dsdna, by Thermo Fisher Scientific (2 mentions) Np4 bsa, by Biosearch Technologies (2 mentions) Np29 bsa, by Biosearch Technologies (2 mentions) Clone 17a2, by BD (1 mentions) Anti cd45r, by BD (1 mentions) Nunc immuno maxisorp plates, by Thermo Fisher Scientific (1 mentions) Np5 bsa, by Biosearch Technologies (1 mentions) Np30 bsa, by Biosearch Technologies (1 mentions) Opteia tmb substrate, by BD (1 mentions) Spectramax 190 microplate reader, by Molecular Devices (1 mentions) 3 3 5 5 tetramethylbenzidine substrate, by Thermo Fisher Scientific (1 mentions) Super aquablue elisa substrate, by Thermo Fisher Scientific (1 mentions) Anti igg3, by Southern Biotech (1 mentions) Np bsa, by Biosearch Technologies (1 mentions) April, by Thermo Fisher Scientific (1 mentions) Milliplex map human autoimmune autoantibody panel haiab 10k, by Merck Group (1 mentions) Multiskan go spectrophotometer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

IgG2c (62 citations) Goat anti-mouse IgG2c-HRP (10 citations) Goat anti-mouse IgG2c (9 citations) Anti-mouse IgG2c (9 citations) Anti-mouse IgG2C-HRP (6 citations) Biotinylated goat-anti-mouse IgG2c (3 citations) Anti-mouse IgG2c (1079-05, (2 citations) Biotinylated anti-mouse IgG2c antibody (2 citations)

8 protocols using anti igg2c

Donor-Specific Antibody Detection

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Balb/c mice (MHC haplotype H2^d) were given an intraperitoneal
injection with 4 × 10⁶ splenocytes from C57 Bl/6 mice (MHC
haplotype H2^b). Plasma from the sensitized Balb/c mice was harvested
21 days later. Flow cross-match was performed using C57 Bl/6 splenocytes as
targets to demonstrate donor antibody specificity as described previously [34 ]. Briefly, C57 Bl/6 splenocytes were
incubated with plasma, visualized with anti-CD3 (BioLegend, clone 17A2),
anti-CD45R (BD Pharmingen, clone RA3–6B2), anti-IgG (BioLegend, clone
Poly4053), anti-IgM (BD Pharmingen, clone R6–60.2), anti-IgG1 (BD
Horizon, clone A85–1), anti-IgG2a (BioLegend, clone RMG2a-62), anti-IgG2b
(R&D Systems, Cat N: F0133), anti-IgG2c (Southern Biotech, Cat N:
1079–02). Cells were gated for singlets and CD3+ cells, then the mean
fluorescence intensity (MFI) was determined for each isotype. Plasma was also
collected from non-sensitized Balb/c mice.

Wilson N.A., Dylewski J., Degner K.R., O’Neill M.A., Reese S.R., Hidalgo L.G., Blaine J, & Panzer S.E. (2019). An in vitro model of antibody-mediated injury to glomerular endothelial cells: upregulation of MHC class II and adhesion molecules. Transplant immunology, 58, 101261.

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Quantifying Antibody Responses via ELISA

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96-well Nunc-Immuno MaxiSorp plates (Thermo Fisher Scientific) were precoated overnight at 4°C with 2 μg/ml anti-IgM, anti-IgG, anti-IgG1, and anti-IgG2C (Southern Biotech) or 50 µg/ml NP(5)-BSA and NP(30)-BSA (Biosearch Technologies). Plates were blocked for 1 h with 2% BSA in PBS before addition of diluted serum (1:6,250, 1:31,250, 1:156,250) for 2 h at RT or overnight at 4°C. Specific antibodies were detected by using goat anti–mouse IgM, IgG, IgG1, IgG3, or rat anti-IgG2C–horseradish peroxidase (1:2,000 dilution; Southern Biotech), and peroxidase reactions were developed by using OptEIA TMB substrate (BD Biosciences) and stopped with sulfuric acid. Absorbance at 450 nm was read by using a SpectraMax 190 microplate reader (Molecular Devices). 450-nm absorbance was corrected by subtraction of 570-nm absorbance.

Wray-Dutra M.N., Chawla R., Thomas K.R., Seymour B.J., Arkatkar T., Sommer K.M., Khim S., Trapnell C., James R.G, & Rawlings D.J. (2018). Activated CARD11 accelerates germinal center kinetics, promoting mTORC1 and terminal differentiation. The Journal of Experimental Medicine, 215(9), 2445-2461.

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ELISA to Assess RBD-specific Antibodies

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An enzyme-linked immunosorbent assay (ELISA) was performed as previously described [26] . Briefly, RBD served as a solid-phase antigen, and the serum obtained from each mouse was serially diluted in duplicate. Immunoenzymatic assays were performed using mouse anti-IgG antibodies conjugated to peroxidase (Sigma-Aldrich, USA). Additionally, IgG subclass responses were assessed using immunized mouse sera treated with anti-IgG1 (Sigma-Aldrich, St. Louis, MO, USA) or anti-IgG2c (SouthernBiotech, Birminghan, USA) antibodies conjugated to peroxidase. To evaluate the strength of antigen-antibody interaction in ELISA, a step involving incubation with increasing concentrations of ammonium thiocyanate (0–4 M), prior to incubation with secondary antibodies (conjugated to peroxidase), was added to the regular protocol.

Rodrigues-Jesus M.J., Teixeira de Pinho Favaro M., Venceslau-Carvalho A.A., de Castro-Amarante M.F., da Silva Almeida B., de Oliveira Silva M., Andreata-Santos R., Gomes Barbosa C., Brito S.C., Freitas-Junior L.H., Boscardin S.B, & de Souza Ferreira L.C. (2022). Nano-multilamellar lipid vesicles promote the induction of SARS-CoV-2 immune responses by a protein-based vaccine formulation. Nanomedicine, 45, 102595.

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Serum Antibody Profiling by ELISA

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Serum was collected by submandibular bleed and microcentrifugation at >18,000 g for 30 min at 4°C. Half area high-binding 96-well plates (Costar) were used for all assays. For total Ig at steady state, plates were coated with goat anti-Ig (2 µg/ml; SouthernBiotech) overnight in coating buffer bicarbonate/carbonate coating buffer (100 mM, pH 9.6). For NP-specific antibody ELISAs, plates were coated with NP-BSA (5 μg/ml; LGC Biosearch Technologies). Diluted serum was added overnight at 4°C or at room temperature for 2 h and detected with biotinylated goat anti-IgG1, anti-IgG3, anti-IgG2b, anti-IgG2c, anti-IgA, or anti-IgM (2 µg/ml; SouthernBiotech) followed by HRP-avidin (1:5,000; SouthernBiotech) and developed with Super AquaBlue ELISA substrate or 3,3′,5,5′-tetramethylbenzidine substrate (Thermo Fisher Scientific) with the manufacturer’s recommended absorbance. For total IgE, the process was the same, except that plates were coated with anti-IgE (2 µg/ml, clone R35-72) and detected with biotinylated anti-IgE (2 µg/ml, clone R35-118).

Wigton E.J., Mikami Y., McMonigle R.J., Castellanos C.A., Wade-Vallance A.K., Zhou S.K., Kageyama R., Litterman A., Roy S., Kitamura D., Dykhuizen E.C., Allen C.D., Hu H., O’Shea J.J, & Ansel K.M. (2021). MicroRNA-directed pathway discovery elucidates an miR-221/222–mediated regulatory circuit in class switch recombination. The Journal of Experimental Medicine, 218(11), e20201422.

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Optimizing Antibody Subtype Detection in ELISAs

Cited 1 time

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For NP-specific ELISAs, plates were coated with NP29-BSA (low affinity) or NP4-BSA (high affinity) (Biosearch Technologies). For ANA ELISAs, plates were coated with poly-L-lysine (Sigma Aldrich) and then subsequently coated with salmon sperm dsDNA (Invitrogen) or Smith ribonucleoprotein (SmRNP) (Arotec Diagnostics). Nucleosome plates were generated by coating with dsDNA followed by histone (Sigma Aldrich). For detection, biotinylated anti-IgG (Jackson Immunoresearch), anti-IgG1, anti-IgG2a, anti-IgG2b, or anti-IgG2c (Southern Biotech), followed by streptavidin-alkaline phosphatase (Vector Laboratories) were used for the detection of respective subtypes. Development was performed with PNPP (p-nitrophenyl phosphate, disodium salt) substrate (Thermo Fisher). Quantitation was performed as previously described (44 (link)).

Schell S.L., Bricker K.N., Fike A.J., Chodisetti S.B., Domeier P.P., Choi N.M., Fasnacht M.J., Luckenbill S.A., Ziegler S.F, & Rahman Z.S. (2021). Context-dependent miR-21 regulation of TLR7-mediated autoimmune and foreign antigen driven antibody-forming cell and germinal center responses. Journal of immunology (Baltimore, Md. : 1950), 206(12), 2803-2818.

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Autoantibody Profiling in Parabiont Pairs

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MILLIPLEX MAP Human Autoimmune Autoantibody Panel (HAIAB-10K, Millipore Sigma) was used following manufacturer’s instructions. Due to supply chain issues, ribosomal P antigen was not included in the original kit, and the assay was performed against 19 autoantigens instead of the original 20. Serum was collected from whole blood by allowing clotting at room temperature for 30 minutes, followed by 2 centrifugation steps at 4°C: 1000g for 10 minutes, and 2700g for 2 minutes, collecting the supernatant after each step. BPI (Surmonics, Cat. A192), Ro60 (Surmonics, Cat.A174), and APRIL (Peprotech, Cat. 310–10C) were added to the assay. For anti-IgG2c (Cat 1079–09, Southern Biotech) and IgG-PE (Cat 1030–09, Southern Biotech), the amount of protein used for coating the beads was 0.005 nmoles; for the other proteins BPI, APRIL, Ro60 the concentration was 0.5 nmoles. For all beads, the calculations were made to have at least 400 beads of each protein (region IDs) at the start of each assay. Serum from 3 parabiont pairs of each type was included in the autoantigen array.

van den Broek T., Oleinika K., Rahmayanti S., Castrillon C., van der Poel C.E, & Carroll M.C. (2023). Invasion of spontaneous germinal centers by naive B cells is rapid and persistent. bioRxiv.

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Quantifying OVA-Specific Antibody Titers

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For OVA-specific IgG1, IgG2b, IgG2c or IgM titres, plasma was serially diluted (4-fold dilutions) and added to ELISA plates (Maxisorp, Nunc, MA USA) pre-coated with 10μg/ml OVA (Sigma Grade V, A5503 MO USA). OVA-specific antibody was detected using biotinylated anti-mouse antibodies (anti-IgG1 [RMG1-1] 0.02μg/ml, anti-IgG2b [RMG2b-1] 0.05μg/ml, Biolegend, CA, USA; anti-IgG2c 0.2μg/ml, Southern Biotech, CA USA; anti-IgM 0.2μgml, Thermo Fisher MA USA) followed by SA-HRP (1:2000 dilution, #P030701-2, Dako, CA USA), and visualised with TMB (#421101, Biolegend, CA USA). Absorbance was measured at 450nm and corrected for absorbance at 540nm using a MultiSkan Go spectrophotometer (Thermofisher, MA USA).

Brooks J.F., Murphy P.R., Barber J., Wells J.W., & Steptoe R.J. (2020). Peripheral tolerance checkpoints imposed by ubiquitous antigen expression limit antigen-specific B-cell responses under strongly immunogenic conditions.

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Quantification of Antibody Subtypes

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Schell S.L., Bricker K.N., Fike A.J., Chodisetti S.B., Domeier P.P., Choi N., Fasnacht M.J., Luckenbill S.A., Ziegler S.F., & Rahman Z.S.M. (2021). Context-dependent miR-21 regulation of TLR7-mediated autoimmune and foreign antigen driven antibody-forming cell and germinal center responses.

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