Mouse igg2a

Manufactured by BioLegend

Sourced in Colombia

Mouse IgG2a is an immunoglobulin G (IgG) subclass found in mice. It is a common laboratory reagent used in various immunological applications.

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Lab products found in correlation

Rat igg2b, by BioLegend (4 mentions) Rat igg2a, by BioLegend (4 mentions) Mouse igg1, by BioLegend (3 mentions) Sheep serum, by Merck Group (2 mentions) Mouse igg2b, by BioLegend (2 mentions) Ifn α, by Merck Group (1 mentions) Recombinant il 12, by R&D Systems (1 mentions) Recombinant ifn α, by Miltenyi Biotec (1 mentions) Biotinylated horse anti mouse igg, by Vector Laboratories (1 mentions) Recombinant human m csf, by R&D Systems (1 mentions) Anti ifnar antibody, by PBL Assay Science (1 mentions) Mouse igg2b, by Merck Group (1 mentions) Ifn γ, by BD (1 mentions) Apc conjugated anti human pd l1 antibody, by BioLegend (1 mentions) Facscalibur, by BD (1 mentions) Facsaria 3 cell sorter, by BD (1 mentions) Dm5500b microscope, by Leica (1 mentions) Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions) Af488 conjugated donkey anti rabbit secondary ab, by Jackson ImmunoResearch (1 mentions) Pe conjugated anti hsp70 clone w27, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

IgG2a (24 citations) Anti-mouse IgG2a (5 citations) Rat IgG2a anti-mouse Mcpt8 antibody (clone TUG8, (4 citations) Biotin-conjugated anti-mouse IgG1 and IgG2a (2 citations) Mouse IgG2a isotype control (2 citations) Mouse IgG2a κ-PE (2 citations) APC mouse IgG2a kappa isotype control (clone MOPC-173 (2 citations) FITC-labeled mouse IgG2a (2 citations) FITC-conjugated mouse IgG2a (2 citations) Biotinylated rat anti-mouse IgG2a (2 citations)

17 protocols using mouse igg2a

Cytokine Stimulation of Immune Cells

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PBMC or NK cells were cultured overnight before treating with purified IFN‐α (Sigma) or recombinant IFN‐α (Miltenyi Biotec), recombinant IL‐12 (R&D systems and Peprotech) or recombinant IL‐15 (Miltenyi Biotec) as described in the text and figure legends. For IFN‐I neutralization, a cocktail of anti‐human interferon α/β receptor chain 2 antibody (clone MMHAR‐2), anti‐human interferon‐α (sheep polyclonal) and anti‐human interferon‐β (sheep polyclonal; all from PBL Assay Science) or a control cocktail of mouse IgG2a (BioLegend) and sheep serum (Sigma) was used, as described previously [25 (link)].

Wantoch M., Wilson E.B., Droop A.P., Phillips S.L., Coffey M., El‐Sherbiny Y.M., Holmes T.D., Melcher A.A., Wetherill L.F, & Cook G.P. (2022). Oncolytic virus treatment differentially affects the CD56dim and CD56bright NK cell subsets in vivo and regulates a spectrum of human NK cell activity. Immunology, 166(1), 104-120.

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Macrophage Differentiation and Stimulation

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Human recombinant M-CSF (R&D Systems) was used for differentiation of blood monocytes into monocyte-derived macrophages (MDMs). Human recombinant IFN-β (BD Biosciences) and IFN-γ (BD Biosciences) were used for macrophage stimulations at the concentrations indicated. Anti-IFNAR antibody (PBL Assay Science) and corresponding isotype antibody mouse IgG2a (BioLegend) were used for neutralization experiments. Immunoperoxidase staining was performed with NUPR1 antibody (Abbexa), corresponding isotype antibody mouse IgG2b (Sigma), CD3 antibody (BD Pharmingen) and Biotinylated horse anti-mouse IgG (Vector).

R. Andrade P., Mehta M., Lu J., M. B. Teles R., Montoya D., O. Scumpia P., Nunes Sarno E., Ochoa M.T., Ma F., Pellegrini M, & Modlin R.L. (2019). The cell fate regulator NUPR1 is induced by Mycobacterium leprae via type I interferon in human leprosy. PLoS Neglected Tropical Diseases, 13(7), e0007589.

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PD-L1 and PD-L2 Expression Analysis

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Cells were stained for PE-conjugated anti-human PD-L2 (Cat#329606, Biolegend) or APC-conjugated anti-human PD-L1 antibody (Cat#329708, Biolegend) at 1:100 dilution on ice for 30 mins and analyzed by BD FACSCalibur. Isotype controls are mouse IgG2a, κ (Cat#400213, Biolegend) and mouse IgG2b, κ (Cat#400322, Biolegend), respectively. GL261 cells overexpressing PD-L1 and/or PD-L2 were stained for PE-conjugated anti-mouse PD-L2 (Cat#107205, Biolegend) and/or APC-conjugated anti-mouse PD-L1 antibody (Cat#124312, Biolegend) at 1:100 dilution on ice for 30 minutes and sorted using a BD FACSAria III cell sorter. Isotype controls were Rat IgG2a, κ (Cat#400508, Biolegend) and Rat IgG2b, κ (Cat#400612, Biolegend), respectively.

Fu Y., Liu C.J., Kobayashi D.K., Johanns T.M., Bowman-Kirigin J.A., Schaettler M.O., Mao D.D., Bender D., Kelley D.G., Uppaluri R., Bi W.L., Dunn I.F., Tao Y., Luo J., Kim A.H, & Dunn G.P. (2020). GATA2 Regulates Constitutive PD-L1 and PD-L2 Expression in Brain Tumors. Scientific Reports, 10, 9027.

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Immunofluorescent Imaging of Tumor Cell Death and HSP70 Expression

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Immunofluorescent imaging was performed as previously described [99 (link)]. Briefly, tumor-bearing and contralateral kidneys were harvested on day 8 of the tumor challenge/therapy scheme described above (1 day post 100μg MD5-1 mAb treatment). Tissues were snap frozen in OCT, cut to 7 μm thickness, and fixed in acetone for 10 min at −20°C. To assess tumor cell death in vivo, sections were stained using unconjugated rabbit anti-cytokeratin 8 (NB100-91850) and 18 (NBP1-67610; Novus Biologics; Littleton, CO), AF488-conjugated donkey anti-rabbit secondary Ab (Jackson ImmunoResearch Laboratories; West Grove, PA), PE-conjugated anti-CD31 (MEC13.3; eBioscience), and AlexaFluor 647-conjugated anti-human/mouse cleaved PARP (Asp214, clone F21-852; BD Pharmingen). To assess HSP70 expression in vivo after Minnelide treatment, sections were stained using unconjugated rabbit anti-cytokeratin 8 (NB100-91850) and 18 (NBP1-67610; Novus Biologics; Littleton, CO), AF488-conjugated donkey anti-rabbit secondary Ab (Jackson ImmunoResearch Laboratories; West Grove, PA), and PE-conjugated anti-HSP70 (clone W27; Santa Cruz Biotechnology) or mouse IgG2a isotype control (BioLegend). Cover slides were applied using ProLong Gold® Antifade Mountant with DAPI (Life Technologies) and imaged with a Leica DM5500 B microscope.

Brincks E.L., Kucaba T.A., James B.R., Murphy K.A., Schwertfeger K.L., Sangwan V., Banerjee S., Saluja A.K, & Griffith T.S. (2015). Triptolide Enhances the Tumoricidal Activity of TRAIL Against Renal Cell Carcinoma. The FEBS journal, 282(24), 4747-4765.

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Measurement of EGFR Inhibition by Immune Sera

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The ability of immune sera to inhibit the growth of the EGFR overexpressing A431 or D17 cell lines were measured using a ³H-thymidine incorporation assay as previously described using 4000 A431 cells/well, 1000 D17 cells/well and 4 × 10⁴ MDA-MB-453 cells/well [6] (link). Control antibodies included cetuximab (10 µg/ml), rituximab (anti-CD20; 10 µg/ml), Ab-1 (10 µg/ml), mouse IgG2a (BioLegend; Dedham, MA) and trastuzumab (21 μg/ml). Percent cell growth inhibition: [(Pre-immune serum CPM – Immune serum CPM)/pre-immune serum CPM] x 100.

Doyle H.A., Gee R.J., Masters T.D., Gee C.R., Booth C.J., Peterson-Roth E., Koski R.A., Helfand S.C., Price L., Bascombe D., Jackson D., Ho R., Post G.R, & Mamula M.J. (2021). Vaccine-induced ErbB (EGFR/HER2)-specific immunity in spontaneous canine cancer. Translational Oncology, 14(11), 101205.

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Comprehensive Flow Cytometry Antibody Panel

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The antibodies for flow cytometry experiments: anti-mouse CD3; anti-mouse CD4; anti-mouse CD8a; anti-mouse CD25; anti-mouse CD38; anti-mouse CD69; anti-mouse CD70; anti-mouse CD11b; anti-mouse CD11c; anti-mouse CD19; anti-CD62L; anti-mouse CD80; anti-mouse CD86; anti-mouse CD68; anti-mouse F4/80; anti-mouse TNFα; anti-Granzyme B; anti-mouse/human CD44; anti-mouse CD183 (CXCR3); anti-mouse CD184 (CXCR4); anti-mouse CD185 (CXCR5); anti-mouse CD186 (CXCR6); anti-mouse CD1d; anti-mouse CD120a (TNFRSF1a); anti-mouse CD152 (CTLA-4); anti-mouse CD279 (PD-1); anti-mouse CD274 (B7-H1 or PD-L1); anti-mouse CD273 (B7-DC or PD-L2); anti-mouse CD117 (c-Kit); anti-mouse Ly-6A/E (SCA-1); anti-mouse Lineage Cocktail with Isotype Ctrl; mouse IgG1, κ Isotype; mouse IgG2b, κ Isotype Ctrl; mouse IgG2a, κ Isotype; rat IgG2b, κ Isotype; Armenian Hamster IgG Isotype; Syrian Hamster IgG Isotype; rat IgG2a, κ Isotype and rat IgG1, κ Isotype Ctrl were procured from BioLegend. The anti-mouse CD34, anti-perforin 1 antibodies and regulatory T cells (Treg) staining kit were purchased from eBioscience.

Kumar S., Singh S.K., Viswakarma N., Sondarva G., Nair R.S., Sethupathi P., Dorman M., Sinha S.C., Hoskins K., Thatcher G., Rana B, & Rana A. (2020). Rationalized inhibition of mixed lineage kinase 3 and CD70 enhances life span and antitumor efficacy of CD8+ T cells. Journal for Immunotherapy of Cancer, 8(2), e000494.

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Quantifying SARS-CoV-2 S1 Protein Binding

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JAWSII cells (ATCC) were seeded onto a 96 well hanging drop plate (Sigma-Aldrich) at 5 x 10⁴ cells/well. EDV only, EDV-αGC, EDV-CONTROL, EDV-COVID and EDV-COVID-αGC were co-incubated with the cells at 1x10⁹ EDVs per well. Untreated JAWSII cells were used as controls. The samples were cultured at 37°C with 5% CO₂ for 48 h before being collected and co-stained with PE anti-mouse αGC:CD1d complex antibody (ThermoFisher, 1:2000) and SARS-CoV-2 S1 protein polyclonal primary antibody (GeneTex, 1:2000) at room temperature for 30 min in the dark. The samples were then stained with Alexa Fluor 647 goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody (ThermoFisher, 1:1000) at 4°C for a further 20 min and analysed using a Gallios flow cytometer (Beckman Coulter). Mouse IgG2a (Cat. #400214, Biolegend) and rabbit IgG (Abcam) were used as isotype controls. DAPI was used to differentiate live/dead cells and single stained samples were used to generate compensation. The samples were analysed using the Kaluza analysis software (V2.1, Beckman Coulter).

Gao S.Y., Amaro-Mugridge N.B., Madrid-Weiss J., Petkovic N., Vanegas N., Visvanathan K., Williams B.R., MacDiarmid J.A, & Brahmbhatt H. (2023). Nanocell COVID-19 vaccine triggers a novel immune response pathway producing high-affinity antibodies which neutralize all variants of concern. Frontiers in Immunology, 13, 1038562.

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Immunophenotyping of Endothelial Cells

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Staining of the surface markers APC-CD31 (BioLegend), PerCP/Cy5.5-CD144 (BioLegend) and mouse IgG2a (BioLegend) was performed in PBS containing 2% fetal bovine serum (v/v) on ice. Flow cytometry data were obtained with a FACSCanto II (BD Biosciences) and analyzed by using FlowJo software (Tree Star).

Guan G., Huo D., Li Y., Zhao X., Li Y., Qin Z., Sun D., Yang G., Yang M., Tan J., Zeng W, & Zhu C. (2021). Engineering hiPSC-CM and hiPSC-EC laden 3D nanofibrous splenic hydrogel for improving cardiac function through revascularization and remuscularization in infarcted heart. Bioactive Materials, 6(12), 4415-4429.

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Quantitative GD2 Expression Analysis

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Formalin-fixed paraffin-embedded tumor sections were immunohistochemically stained with mouse anti-human GD2 antibody (BD Biosciences, Cat# 554,272, 0.5 mg/mL). The isotype control was stained with purified mouse IgG2a (BioLegend, Cat# 400,202). All images were captured from tumor sections using a Zeiss Axio Vert.A1 microscope and Zeiss ZEN imaging software. The tissue staining intensity was compared with that from positive and negative controls and scored from 0 to 4 according to two components: staining intensity and the percentage of positive cells. Each sample was assessed and graded by two independent observers.

Yu L., Huang L., Lin D., Lai X., Wu L., Liao X., Liu J., Zeng Y., Liang L., Zhang G., Wang B., Wu Z., Tao S., Liu Y., Jiao C., Chang L.J, & Yang L. (2021). GD2-specific chimeric antigen receptor-modified T cells for the treatment of refractory and/or recurrent neuroblastoma in pediatric patients. Journal of Cancer Research and Clinical Oncology, 148(10), 2643-2652.

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Exosome Surface Marker Characterization

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Antibodies to CD9 (HI9a), CD63 (NVG-2), CD63 (H5C6), CD81 (Eat-2), CCR9 (9B1), CXCR4 (L276F12), and CX3CR1 (SA011F11) were purchased from BioLegend (San Diego, CA). Isotype controls including Rat IgG2a, Rat IgG2b, Armenian Hamster IgG, Mouse IgG2a, and Mouse IgG1 were also from BioLegend. The antibody to CD9 (KMC8) was obtained from BD Biosciences. The antibodies to CCR7 (4B12), CCR10 (248918) were acquired from R&D Systems. The cells or microbead-conjugated exosomes were stained with the fluorescently labeled antibodies, washed twice with PBS containing 2% FBS and 2 mM ethylenediaminetetraacetic acid (EDTA) (Wako, Osaka, Japan), and analyzed by using BD Accuri C6 flow cytometer and software (BD Biosciences). For this method of microbead conjugation of exosomes, only positive events can be detected and fluorescently quantified in the flow cytometry. In some experiments, total (intracellular plus surface) staining experiment was done by using FIX and PERM Kit (Thermo Fisher Scientific) according to manufacturer's instructions.

Park E.J., Myint P.K., Appiah M.G., Worawattananutai P., Inprasit J., Prajuabjinda O., Soe Z.Y., Gaowa A., Kawamoto E, & Shimaoka M. (2021). Ligand-competent fractalkine receptor is expressed on exosomes. Biochemistry and Biophysics Reports, 26, 100932.

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