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Human igg elisa kit

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The Human IgG ELISA Kit is a laboratory tool used to quantitatively measure the levels of human immunoglobulin G (IgG) in a sample. It is a standardized enzyme-linked immunosorbent assay (ELISA) that provides a reliable and efficient method for the detection and analysis of IgG antibodies.

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Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Mouse anti dsdna igg elisa kit, by Alpha Diagnostic (2 mentions) Complement 3 elisa kit, by Alpha Diagnostic (2 mentions) Balb canncrlcrlj mice, by Charles River Laboratories (1 mentions) Isostrip mouse monoclonal antibody isotyping kit, by Roche (1 mentions) Human tnf elisa kit, by BD (1 mentions) Eti ab auk plus, by DiaSorin (1 mentions) Biolux gaussia luciferase assay kit, by New England Biolabs (1 mentions) Protein a sepharose, by GE Healthcare (1 mentions) Human iga elisa kits, by Fortis Life Sciences (1 mentions)

10 protocols using human igg elisa kit

1

Generation of Anti-Insoluble Fibrin mAbs

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To produce new anti-insoluble fibrin mAbs, 6-week-old BALB/cAnNCrlCrlj mice (Charles River Japan, Yokohama, Japan) or rats were immunized intraperitoneally with an emulsion of Freund's complete adjuvant (DIFCO, New Jersey, USA) and a saline solution containing 50 μg of the epitope peptide from the fibrinogen β chain that we previously discovered [7 (link)]. Three to five successive booster injections were administered intraperitoneally at 2-week intervals using the same amount of antigen in an adjuvant system (Merck, Hessen, Germany). A final boost was provided by administering the same amount of antigen intravenously. Animals were sacrificed under anesthesia, and the iliac lymph node from rats or spleen from mice was extracted and cells were fused to the P3X63-Ag8.653 myeloma cell line. Then, primary hybridoma cells were selected with HAT medium and cloned by limited dilution. Hybridoma cells were acclimated to appropriate serum-free media for large scale culture. Supernatant titer was confirmed using the Human IgG ELISA Kit (Bethyl Laboratories, Massachusetts, USA) and mAbs were obtained by affinity purification and size exclusion chromatography. The isotypes of the mAbs were determined using IsoStrip Mouse Monoclonal Antibody Isotyping Kit (Roche, Basel-Stadt, Switzerland).
Fuchigami H, & Matsumura Y. (2022). Characterization of antibody clones that bind exclusively to insoluble fibrin. Blood Coagulation & Fibrinolysis, 34(1), 20-27.
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2

Engrafting Human MM Samples in NOG Mice

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Human MM samples were obtained from patients at Tokyo Medical and Dental University or National Cancer Center Hospital. All patients provided informed consent, and the study was approved by the Institutional Review Board. For primary MM samples obtained from iliac bone marrow, mononuclear cells were purified using a Ficoll‐density gradient centrifugation enrichment protocol and transplanted into 6‐week‐old NOD.Cg‐Prkdcscid Il2rgtm1Sug/Jic (NOG) mice (Central Institute for Experimental Animals, Kawasaki, Japan) by tibial intramedullary injection. Successful engraftment was confirmed by detecting human immunoglobulins in the serum of mice using ELISA (Human IgG ELISA Kit; Bethyl Laboratories, Montgomery, TX, USA), and their levels were measured monthly to evaluate the drug response. Mice were killed when treatment was completed and bone marrow cells were collected for flow cytometric analysis.
Nakagawa M., Fujita S., Katsumoto T., Yamagata K., Ogawara Y., Hattori A., Kagiyama Y., Honma D., Araki K., Inoue T., Kato A., Inaki K., Wada C., Ono Y., Yamamoto M., Miura O., Nakashima Y, & Kitabayashi I. (2018). Dual inhibition of enhancer of zeste homolog 1/2 overactivates WNT signaling to deplete cancer stem cells in multiple myeloma. Cancer Science, 110(1), 194-208.
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3

Quantifying Autoantibodies and Complement in Mice

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Mouse serum was diluted 1:100 with assay buffer for anti-dsDNA antibody detection and 1:25000 for complement 3 detections. Assays were carried out with Mouse Anti-dsDNA IgG ELISA Kit (5120, Alpha Diagnostic Intl. Inc.) and Complement 3 ELISA Kit (6270, Alpha Diagnostic Intl. Inc.), following the manufacturer’s protocols. Urine protein concentrations were detected using BCA Protein Assay Kit (23225, Thermo Scientific). IgG of B cell cultural supernatant was measured by Human IgG ELISA Kit (E88–104, Bethyl).
Li P., Jiang M., Li K., Li H., Zhou Y., Xiao X., Xu Y., Krishfield S., Lipsky P.E., Tsokos G.C, & Zhang X. (2021). Glutathione peroxidase 4 regulated neutrophil ferroptosis induces systemic autoimmunity. Nature immunology, 22(9), 1107-1117.
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4

Quantifying Autoantibodies and Complement in Mice

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Mouse serum was diluted 1:100 with assay buffer for anti-dsDNA antibody detection and 1:25000 for complement 3 detections. Assays were carried out with Mouse Anti-dsDNA IgG ELISA Kit (5120, Alpha Diagnostic Intl. Inc.) and Complement 3 ELISA Kit (6270, Alpha Diagnostic Intl. Inc.), following the manufacturer’s protocols. Urine protein concentrations were detected using BCA Protein Assay Kit (23225, Thermo Scientific). IgG of B cell cultural supernatant was measured by Human IgG ELISA Kit (E88–104, Bethyl).
Li P., Jiang M., Li K., Li H., Zhou Y., Xiao X., Xu Y., Krishfield S., Lipsky P.E., Tsokos G.C, & Zhang X. (2021). Glutathione peroxidase 4 regulated neutrophil ferroptosis induces systemic autoimmunity. Nature immunology, 22(9), 1107-1117.
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5

Pharmacokinetics of R2agoTNF in Mice

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Male BALB/c mice (8-weeks-old) were injected i.p. with 30 μg R2agoTNF or scR2agoTNF-Fc. To investigate pharmacokinetics, blood was collected from the tail vein of mice at an appropriate interval after injection. Protein concentrations in plasma samples were measured by a human TNF ELISA kit (BD Biosciences) or human IgG ELISA kit (Bethyl Laboratories, Montgomery, TX, USA). The t½ and AUC were calculated by moment analysis.
Inoue M., Tsuji Y., Ueno R., Miyamoto D., Tanaka K., Moriyasu Y., Shibata S., Okuda M., Ando D., Abe Y., Kamada H, & Tsunoda S.I. (2023). Bivalent structure of a TNFR2-selective and agonistic TNF-α mutein Fc-fusion protein enhances the expansion activity of regulatory T cells. Scientific Reports, 13, 13762.
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6

IgG Antibody Quantification Protocol

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Blood samples were stored overnight at 4°C to coagulate and then spun in a benchtop centrifuge at 1500 ×g for 5 minutes. Serum was collected, transferred into fresh tubes, and then frozen at −80°C for storage. Total IgG and neutralizing antibody levels were determined by using Human IgG ELISA Kit (Bethyl Laboratories, Montgomery, TX) and ETI-AB-AUK PLUS (DiaSorin, Saluggia, Italy), respectively. IgG subclasses were measured by using human IgG subclass kits (Invitrogen). Each sample was run in duplicate and a standard curve was included in each plate.
Ma L., Norton M.G., Mahmood I., Zhao Z., Zhong L., Zhang P, & Struble E.B. (2014). Transplacental Transfer of Hepatitis B Neutralizing Antibody during Pregnancy in an Animal Model: Implications for Newborn and Maternal Health. Hepatitis Research and Treatment, 2014, 159206.
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7

Quantifying Palivizumab and Gaussia Luciferase in Biological Samples

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Palivizumab levels in the cell culture media, serum and BALF were measured using Human IgG ELISA kit (Bethyl, Cambridge, UK) according to the manufacturer’s instructions. GLux activity in the serum was measured using BioLux Gaussia Luciferase Assay Kit (NEB, Ipswich, USA) according to the manufacturer’s instructions. Expression levels in ELF were corrected for the dilution using lavage fluid collection urea assay as described previously8 (link).
Antepowicz A., Habib O., Kirsebom F., Johansson C., Gill D.R, & Hyde S.C. (2021). Lentiviral and AAV-mediated expression of palivizumab offer protection against Respiratory Syncytial Virus infection. Scientific Reports, 11, 15694.
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8

Cytokine-activated T Cell Co-culture with B Cells

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T cells were activated and treated with cytokines as described earlier. After two (for total CD4+ T cells) or four (for naïve CD4+ T cells) days of activation, cells were either sorted on the basis of forward scatter (FSC) and side scatter (SSC) on a FACS ARIA II cell sorter at the NYU School of Medicine Flow Cytometry Core Facility based on large size or, alternatively, anti-CD3/28 magnetic Dynabeads were removed from cell culture by two rounds of magnetic separation. The cells were washed, counted, and then combined with thawed B cells from the same donor in a 96-well U-bottom plate. Each well contained 2.5 × 103 T cells and 4 × 104 B cells. Low Endotoxin Staphylococcal Enterotoxin B (SEB)(Toxin Technologies) was used final concentration: 0.05 ng/ml in a total of 200 µl/well. After 7 or 14 days, the contents of the wells were centrifuged and the supernatants were analyzed for total IgG concentrations with a Human IgG ELISA Kit (#E88-104, Bethyl Laboratories).
Weiss J.M., Chen W., Nyuydzefe M.S., Trzeciak A., Flynn R., Tonra J.R., Marusic S., Blazar B.R., Waksal S.D, & Zanin-Zhorov A. (2016). ROCK2 signaling is required to induce a subset of T follicular helper cells through opposing effects on STATs in autoimmune settings. Science signaling, 9(437), ra73.
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9

Generation and Characterization of LAIR-2-Fc Fusion Protein

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The recombinant human LAIR-2-Fc protein construct was generated by genetically linking the human LAIR-2 to both ends of the Fc portion of human IgG1. The Fc domain was mutated with N297A to prevent binding to Fc receptors as described previously,24 (link) and were further mutated with T250Q/M428L mutations to increase the binding ability to neonatal FcR (FcRn).25 (link) Anti-CD20 IgG1 (Rituximab; accession number: DB00073) introduced with the same N297A and T250Q/M428L mutations in its Fc portion served as the control. The recombinant protein was expressed transiently in suspended human embryonic kidney (HEK293F) cells. Supernatants were harvested 7 days post-transfection and purified by affinity chromatography using Protein A-SepharoseTM (GE Healthcare). The purity of each recombinant antibody was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Pharmacokinetic (PK) parameters were examined in NOD/SCID mice by injecting LAIR-2-Fc protein (n = 4) at 10 mg/kg, followed by determining the serum antibody concentrations at various time points using human IgG ELISA Kits (Bethyl Laboratories). PK parameters were calculated from the final dataset using PK solver software.
Xu L., Wang S., Li J., Li J, & Li B. (2020). Cancer immunotherapy based on blocking immune suppression mediated by an immune modulator LAIR-1. Oncoimmunology, 9(1), 1740477.
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10

Pharmacokinetics of IgG, IgA Antibodies

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IgG1 CD20, Ig1/IgA2 CD20, or IgA2 CD20 (100 μg/mouse) was injected intravenously into C57BL/6 mice (four mice/group). Blood was collected via the tail vein from alternating mice at the indicated time points. The human IgA antibody concentrations in the sera were determined by enzyme-linked immunosorbent assays (ELISAs) using human IgA ELISA kits (Bethyl Laboratories), and human IgG and IgGA antibody concentrations in the sera were determined using human IgG ELISA Kits (Bethyl Laboratories) according to the manufacturer's instructions. The data were analyzed by PK solver software, as previously described [2 (link)].
Li B., Xu L., Tao F., Xie K., Wu Z., Li Y., Li J., Chen K., Pi C., Mendelsohn A., Larrick J.W., Gu H, & Fang J. (2017). Simultaneous exposure to FcγR and FcαR on monocytes and macrophages enhances antitumor activity in vivo. Oncotarget, 8(24), 39356-39366.
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