Igg2b

Manufactured by Santa Cruz Biotechnology

Sourced in United States

IgG2b is an immunoglobulin isotype that is a part of the IgG class of antibodies. It is a key component in the immune response and plays a crucial role in the adaptive immune system.

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Lab products found in correlation

Igg2a, by Santa Cruz Biotechnology (4 mentions) Igg2c, by Southern Biotech (2 mentions) Multiskan fc, by Thermo Fisher Scientific (2 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Merck Group (1 mentions) Microplate reader, by Bio-Rad (1 mentions) Anti mouse igg, by Merck Group (1 mentions) Stop reagent, by Merck Group (1 mentions) Horseradish peroxidase conjugated goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions) Maxisorp, by Thermo Fisher Scientific (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Goat anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions) Phalloidin ifluor 555 reagent, by Abcam (1 mentions) Sephadex g 150, by Yuanye Bio-Technology (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) Cell counting kit 8 cck 8, by Beyotime (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Qiagen (1 mentions) 96 well plate, by Corning (1 mentions) Igg2c, by Merck Group (1 mentions)

8 protocols using igg2b

Antibody Titers Quantification by ELISA

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The titers of IgG antibodies were measured in the serum sample from each group of mice by an enzyme-linked immunosorbent assay (ELISA). Briefly, the recombinant GP and VP40 proteins were individually coated as the antigens at a concentration of 1 μg/ml in 96-well polystyrene microtiter plates for overnight at 4°C. After blocking with 5% milk in PBS buffer, the ELISA plates were incubated with serial dilutions of serum samples at 37°C for 2 h. The titer of total IgG was detected with horseradish peroxidase (HRP)-conjugated goat against mouse IgG (1:2000 dilution, Santa Cruz Biotechnology). IgG subtype antibodies including IgG1, IgG2a, and IgG2b (1:2000 dilution, Santa Cruz Biotechnology) were further detected individually. The immune complex was developed with 3,3′,5,5′-tetramethylbenzidine (TMB) (Sigma). The reaction was stopped with 2 mol/L H₂SO₄, and the plates were read with the absorbance at 450 nm in a microplate reader (Bio-Rad). All samples were detected in triplicate wells.

Ren S., Wei Q., Cai L., Yang X., Xing C., Tan F., Leavenworth J.W., Liang S, & Liu W. (2018). Alphavirus Replicon DNA Vectors Expressing Ebola GP and VP40 Antigens Induce Humoral and Cellular Immune Responses in Mice. Frontiers in Microbiology, 8, 2662.

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Serum Antibody ELISA for Mtb Antigen

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Serum antibody ELISA was carried out as described previously [83] (link), using 5 µg ml⁻¹ of Mtb antigen for coating and HRP-conjugated anti-mouse IgG (Sigma-Aldrich), IgG1, IgG2a (BD-Pharmingen) and IgG2b (Santa Cruz Biotech) antibodies for detection.

Nandakumar S., Kannanganat S., Posey J.E., Amara R.R, & Sable S.B. (2014). Attrition of T-Cell Functions and Simultaneous Upregulation of Inhibitory Markers Correspond with the Waning of BCG-Induced Protection against Tuberculosis in Mice. PLoS ONE, 9(11), e113951.

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Chikungunya Virus Antibody Titers

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Antibody titers were determined using a virion-based enzyme-linked immunosorbent assay (ELISA) as previously described (39 (link)). CHIKV-coated (10⁶ virions/well in 50 ml of PBS) polystyrene 96-well MaxiSorp plates (Nunc) were blocked with PBS containing 0.05% Tween 20 (PBST) and 5% (wt/vol) nonfat milk for 1.5 h at 37°C. Sera from normal or infected groups were heat inactivated and serially diluted in antibody diluent (0.05% PBST plus 2.5% [wt/vol] nonfat milk). One hundred microliters of diluted sera per well was added, and the mixture was incubate for 1 h at 37°C. Horseradish peroxidase-conjugated goat anti-mouse IgG (Santa Cruz, catalog no. sc-2005), IgG1 (Santa Cruz, catalog no. sc-2060), IgG2b (Santa Cruz, catalog no. sc-2062), IgG2c (Southern Biotech, catalog no. 1079-01), IgG3 (Santa Cruz, catalog no. sc-2972), and IgM (Santa Cruz, catalog no. sc-2973) antibodies were used. IgG2c was tested in lieu of IgG2a since only IgG2c gene is present in C57BL/6 mice (39 (link)). Total IgG and IgM quantification assays were performed on serum samples from animals receiving serum dilutions of 1:2,000 and 1:100, respectively. Pooled sera were used for IgG1, IgG2b, IgG2c, and IgG3 isotyping. ELISAs were developed using TMB substrate and terminated with stop reagent (Sigma-Aldrich). The absorbance was measured at 450 nm.

Teo T.H., Her Z., Tan J.J., Lum F.M., Lee W.W., Chan Y.H., Ong R.Y., Kam Y.W., Leparc-Goffart I., Gallian P., Rénia L., de Lamballerie X, & Ng L.F. (2015). Caribbean and La Réunion Chikungunya Virus Isolates Differ in Their Capacity To Induce Proinflammatory Th1 and NK Cell Responses and Acute Joint Pathology. Journal of Virology, 89(15), 7955-7969.

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Immunomodulatory Effects of Eucommia ulmoides

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Eucommia ulmoides Oliver leaf was obtained from SPH Zunyi Pharmaceutical Co., Ltd. (Zunyi, China). OVA and Freund’s Adjuvant, Complete, was purchased from Sigma-Aldrich Co., LLC (St Louis, MO, USA). EDC and ADH were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Sephadex G-150 was obtained from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Cell Counting Kit-8 (CCK-8) was purchased from Shanghai Beyotime Biotechnology Co., Ltd. (Shanghai, China). The mouse macrophage (RAW264.7) cell line was obtained from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China). Phalloidin-iFluor 555 Reagent was obtained from Abcam Inc. (Abcam, Cambridge, MA, USA) and 4′,6-diamidino-2-phenylindole (DAPI) was obtained from Solarbio (Beijing, China). HRP goat anti-mouse IgG, IgG1, IgG2a, and IgG2b were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). IL-2, IL-4, IL-6, and an IFN-γ mouse uncoated enzyme-linked immunosorbent assay (ELISA) kit were obtained from Thermo Fisher Scientific Inc. (Asheville, NC, USA).

Feng H., Yang J., Zhi H., Hu X., Yang Y., Zhang L., Liu Q., Feng Y., Wu D, & Li H. (2021). Eucommia ulmoides Leaf Polysaccharide in Conjugation with Ovalbumin Act as Delivery System Can Improve Immune Response. Pharmaceutics, 13(9), 1384.

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Antibody Levels Estimation in Mice

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Antibody levels in the mouse serum were estimated using the nApa (2 μg/ml) or WCL (5 μg/ml) for coating and the anti-mouse IgG1, IgG2a (BD-Pharmingen) and IgG2b (Santa Cruz Biotechnology, Dallas, TX) HRP-conjugated antibodies for detection.

Nandakumar S., Kannanganat S., Dobos K.M., Lucas M., Spencer J.S., Amara R.R., Plikaytis B.B., Posey J.E, & Sable S.B. (2016). Boosting BCG-primed responses with a subunit Apa vaccine during the waning phase improves immunity and imparts protection against Mycobacterium tuberculosis. Scientific Reports, 6, 25837.

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ELISA for Antibody Subtyping

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ELISA was performed as described previously (28 (link)). Briefly, 96-well plates (Costar) were coated with 1.2 μg of synthetic SP (>90% purity; Sangon Biotech, Shanghai, China) per well and incubated overnight at 4°C. Individual sera were serially diluted from 1:100 to 1:4,096 in a blocking buffer (0.05% Tween, 3% milk in PBS) and incubated for 1 h at 37°C. Subsequently, 100 μl of horseradish peroxidase (HRP) conjugated rabbit anti-mouse antibody subtypes including IgA, IgG, IgG1, IgG2a, IgG2b, or IgG3 (Santa Cruz) at dilutions of 1:2,500 were added, followed by incubation for 1 h at 37°C. The color reaction was developed with TMB (Qiagen) for 30 min and then stopped with 2 M H₂SO₄. The absorbance was detected at 450 nm. Sera from PBS- and FD^mut-treated mice were used as control. Antibody titers were defined as logarithm₁₀ of the reciprocal of the highest dilution giving twice the OD of negative control sera (29 (link), 30 (link)).

Wan Y., Zhang Y., Wang G., Mwangi P.M., Cai H, & Li R. (2020). Recombinant KRAS G12D Protein Vaccines Elicit Significant Anti-Tumor Effects in Mouse CT26 Tumor Models. Frontiers in Oncology, 10, 1326.

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ZIKV Antibody Titration by ELISA

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Antibody titres were determined by virion‐based ELISA as previously described.¹⁸, ¹⁹ Pooled heat‐inactivated mouse sera of respective groups were tested at 1:100 dilution for IgM, and 1:500 dilution for IgG. For IgG subtyping studies, pooled sera were serially diluted from 1:250 to 1:32 000 for IgG3, 1:512 000 for IgG1 and IgG2b, and 1:8 192 000 for IgG2c. ZIKV‐specific IgG subtypes are expressed as antibody titre, defined as the greatest reacting dilution before the OD value reaches baseline control (pooled 0 dpi sera). HRP‐conjugated goat anti‐mouse IgM (Santa Cruz Biotechnology, Dallas, TX, USA), IgG (Merck, Kenilworth, NJ, USA), IgG1 (Santa Cruz Biotechnology), IgG2b (Santa Cruz Biotechnology), IgG2c (Southern Biotech, Birmingham, AL, USA) and IgG3 (Santa Cruz Biotechnology) antibodies were used. IgG2c was tested in lieu of IgG2a since only IgG2c gene is present in C57BL/6 mice.⁴⁷, ⁶⁷ ELISAs were developed using TMB substrate and terminated with stop reagents (Sigma‐Aldrich). The absorbance was measured at 450 nm. ELISA readings were conducted in duplicates.

Lee C.Y., Carissimo G., Chen Z., Lum F., Abu Bakar F., Rajarethinam R., Teo T., Torres‐Ruesta A., Renia L, & Ng L.F. (2020). Type I interferon shapes the quantity and quality of the anti‐Zika virus antibody response. Clinical & Translational Immunology, 9(4), e1126.

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Quantification of Anti-LG3 Antibodies

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Anti‐LG3 titers were measured by ELISA. Recombinant LG3 without His8G‐tag (1 μg/well) was first coated on 96‐well Immulon II HB plates (Thermo Electron, Waltham, MA). Sera were diluted (1/100) and 100 μl/well were added. After washing, the LG3‐bound mouse Igs were detected using sheep antimouse IgG (Amersham, Baie d'Urfé, QC, Canada), goat antimouse IgG1, IgG2a, IgG2b, IgG3, or goat antimouse IgM (all from Santa Cruz Biotechnology, Santa Cruz, CA) horseradish peroxidase (HRP) conjugate. Reactions were revealed with tetramethylbenzidine substrate (BD Biosciences) and stopped with H₂SO₄ (1 mol/L). Optical densities (OD) were read using a microplate photometer at 450 nm (Multiskan FC; Thermo Fisher Scientific, Nepean, ON, Canada).
Total IgG and antinuclear antibodies (ANA) levels were assessed using Mouse IgG total Ready‐SET‐Go Kits (Affymetrix eBioscience, Santa Clara, CA) and ANA Mouse Bioassay Kits (US Biological Life Sciences, Marblehead, MA), respectively, according to the manufacturer's instructions.

Padet L., Dieudé M., Karakeussian‐Rimbaud A., Yang B., Turgeon J., Cailhier J., Cardinal H, & Hébert M. (2018). New insights into immune mechanisms of antiperlecan/LG3 antibody production: Importance of T cells and innate B1 cells. American Journal of Transplantation, 19(3), 699-712.

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