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Mouse igg isotype control

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The Mouse IgG isotype control is a laboratory reagent used in flow cytometry and immunoassays as a negative control. It is an immunoglobulin G (IgG) antibody from mouse that does not recognize any known antigen, providing a baseline measurement for non-specific binding.

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Lab products found in correlation

αvβ8 blocking adwa16 antibody, by BioXCell (2 mentions) Luciferase assay system kit, by Promega (2 mentions) Vybrant cell labeling solution, by Thermo Fisher Scientific (1 mentions) Anti pd 1 ab, by BioXCell (1 mentions) Rnase, by Thermo Fisher Scientific (1 mentions) Heat inactivated fetal bovine serum, by Thermo Fisher Scientific (1 mentions) Dnase, by Thermo Fisher Scientific (1 mentions) Ht 29, by Thermo Fisher Scientific (1 mentions) Mccoy s 5a medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Doxorubicin, by LC Laboratories (1 mentions) Mcf 7, by American Type Culture Collection (1 mentions) Mel 888, by American Type Culture Collection (1 mentions) Mel 526, by American Type Culture Collection (1 mentions)

5 protocols using mouse igg isotype control

1

Quantifying TGFβ Activation via Luciferase

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Transformed mink lung epithelial cells stably expressing a luciferase construct under the control of a TGFβ-responsive promoter21 (link) (a kind gift from Prof. Dan Rifkin, NYU, New York, NY, USA) were co-cultured with moDC plus either 200 µg ml−1 αvβ8-blocking ADWA16 antibody or 200 µg ml−1 mouse IgG isotype control (BioXcell). Cells were incubated overnight at 37 °C, and luciferase levels measured using the Luciferase assay system kit according to the manufacturer’s instructions (Promega, Madison, WI, USA). An active TGFβ standard curve was used to calculate levels of active TGFβ from luminescence intensity observed.
Fenton T., Kelly A., Shuttleworth E., Smedley C., Atakilit A., Powrie F., Campbell S., Nishimura S., Sheppard D., Levison S., Worthington J., Lehtinen M, & Travis M. (2016). Inflammatory cues enhance TGFβ activation by distinct subsets of human intestinal dendritic cells via integrin αvβ8. Mucosal immunology, 10(3), 624-634.
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2

Phagocytosis of Tumor Cells by DCs

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Dendritic cells were plated in 24-well plates (105 cells/well). After 48 h, lung tumor cells treated with gefitinib at their specific IC50 (see Table 1) or DMSO carrier were labeled with DiO cell-labeling solution (Vybrant Cell-Labeling Solution, Molecular Probes) and added to dendritic cells at a 1:1 ratio. Where indicated, tumor cells were incubated with anti-mouse/human/rat CD47 mAb (10 μg/ml, Bio X Cell) or mouse IgG isotype control (10 μg/ml, Bio X Cell) prior to culture with dendritic cells. Following 2.5 h co-culture at 37°C, cells were washed twice with PBS and then labeled with anti-CD11c mAbs (1:200, Miltenyi). Phagocytosis was determined by flow cytometry detection of dendritic cells double positive for CD11c and DiO cell-labeling solution.
Nigro A., Ricciardi L., Salvato I., Sabbatino F., Vitale M., Crescenzi M.A., Montico B., Triggiani M., Pepe S., Stellato C., Casolaro V, & Dal Col J. (2020). Enhanced Expression of CD47 Is Associated With Off-Target Resistance to Tyrosine Kinase Inhibitor Gefitinib in NSCLC. Frontiers in Immunology, 10, 3135.
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3

Quantifying TGFβ Activation via Luciferase

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Transformed mink lung epithelial cells stably expressing a luciferase construct under the control of a TGFβ-responsive promoter21 (link) (a kind gift from Prof. Dan Rifkin, NYU, New York, NY, USA) were co-cultured with moDC plus either 200 µg ml−1 αvβ8-blocking ADWA16 antibody or 200 µg ml−1 mouse IgG isotype control (BioXcell). Cells were incubated overnight at 37 °C, and luciferase levels measured using the Luciferase assay system kit according to the manufacturer’s instructions (Promega, Madison, WI, USA). An active TGFβ standard curve was used to calculate levels of active TGFβ from luminescence intensity observed.
Fenton T., Kelly A., Shuttleworth E., Smedley C., Atakilit A., Powrie F., Campbell S., Nishimura S., Sheppard D., Levison S., Worthington J., Lehtinen M, & Travis M. (2016). Inflammatory cues enhance TGFβ activation by distinct subsets of human intestinal dendritic cells via integrin αvβ8. Mucosal immunology, 10(3), 624-634.
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4

Anti-PD-1 and ATRA in Tumor Growth

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All animal experiments were performed in accordance with a protocol approved by the Wake Forest Institutional Animal Care and Use Committee. Wild type C57/B6 mice (7–8 weeks) were injected with 2 × 105 LL2 or CMT167 cells suspended in growth medium with 50% of matrigel into the flank region. One week after inoculation, mice were treated with 100μg/mouse anti-PD-1Ab (BioXcell), 100μg/mouse IgG isotype control (BioXcell), 20mg/kg ATRA (BioHems) or combination treatment twice a week. The growth of tumor was measured by Tumor volumes were measured twice a week and calculated by using the formula: Tumor volume=length x width2/2.
Wang Y., Smith M., Ruiz J., Liu Y., Kucera G.L., Topaloglu U., Chan M.D., Li W., Su J, & Xing F. (2023). Modulation of oxidative phosphorylation and mitochondrial biogenesis by cigarette smoke influence the response to immune therapy in NSCLC patients. Lung cancer (Amsterdam, Netherlands), 178, 37-46.
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5

Cancer Cell Line Characterization and Treatment

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Human breast cancer cell lines MCF7 and MDA-MB-468, human melanoma cancer cell lines MEL-526 and MEL-888, human lung carcinoma cell line A549, human prostate cancer cell line PC3, and human colorectal adenocarcinoma cell line HT-29 were purchased from American Type Culture Collection (ATCC). MEL-526, MEL-888, and PC3 cells were cultured in RPMI-1640 (Hyclone). MCF7, MDA-MB-468, and A549 cells were cultured in Dulbecco’s Modified Eagle Medium (Hyclone). HT-29 was cultured in McCoy’s 5A Medium (Gibco). All cells were cultured with presence of 1X penicillin-streptomycin and supplemented with 10% heat-inactivated fetal bovine serum (Gibco) during normal growth conditions. Cells were cultured with antibiotic-free growth medium during drug treatment. Doxorubicin purchased from LC laboratories (#D-4000), anti-mouse PD-1 (#BE0033-2) and mouse IgG isotype control (#BE0086) from BioXCell. The PAK inhibitor PF-03758309 (PAKi) purchased from Selleckchem (#S7094). In nucleic acid depletion assay cells were treated with 10 µ/mL of DNase or 10 µ/mL of RNase (Invitrogen).
Wang Y., Pattarayan D., Huang H., Zhao Y., Li S., Wang Y., Zhang M., Li S, & Yang D. (2024). Systematic investigation of chemo-immunotherapy synergism to shift anti-PD-1 resistance in cancer. Nature Communications, 15, 3178.
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