Tamoxifen (Sigma) was dissolved in corn oil (Sigma) and administered intraperitoneally at 200 mg/kg per day x 3-5 days for lineage labelling and gene deleting studies. For viral infection, adenoviruses (WC-U of Iowa-272) were suspended in PBS and administered intranasally at 1X108 or 1X109 PFU/animal. For depleting Gli1+ cells in vivo, diphtheria toxin (Sigma) was administered intraperitoneally at 50 ng/g of mouse per day x 5 days. For FTY720 treatment, FTY720 was administered intraperitoneally at 3 mg/kg of mouse every two day for 7 doses. For blocking IFNγ in mice, anti-mouse IFNγ (XMG1.2, BioXCell) or IgG isotype control (BioXCell) were administered intranasally at 100 μg/dose weekly per animal for 7 doses. For depletion of T cells in vivo, anti-mouse IL7R (BioXCell) or IgG isotype control (BioXCell) were administered intranasally at 100 μg/dose weekly per animal for 7 doses. At indicated time points, lungs were collected for flow cytometry or histological analysis.
Igg isotype control
Manufactured by BioXCell
Sourced in United States
The IgG isotype control is a laboratory reagent used in flow cytometry and related immunoassays. It serves as a negative control to establish background signal levels, enabling more accurate data interpretation.
Automatically generated - may contain errors
Lab products found in correlation
Anti pd 1 checkpoint inhibitor antibodies, by BioXCell (2 mentions)
Etoposide, by Accord Healthcare (2 mentions)
Azd1775, by Selleck Chemicals (2 mentions)
Anti pd l1 antibodiy clone 10f 9g2, by BioXCell (2 mentions)
C 176, by Selleck Chemicals (2 mentions)
Recombinant ifn γ, by BioLegend (2 mentions)
Cisplatin, by Accord Healthcare (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Anti pd 1 antibody, by BioXCell (2 mentions)
Tamoxifen, by Merck Group (1 mentions)
Diphtheria toxin, by Merck Group (1 mentions)
Corn oil, by Merck Group (1 mentions)
Anti mouse ifn γ xmg1, by BioXCell (1 mentions)
Isotype control igg, by R&D Systems (1 mentions)
Anti cd8 antibody, by BioXCell (1 mentions)
Mda mb 231, by Thermo Fisher Scientific (1 mentions)
Bt 549, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Cd45 apc cy7, by BioLegend (1 mentions)
Cd4 fitc, by BioLegend (1 mentions)
25 protocols using igg isotype control
1
Targeted Lineage Labeling and Cell Depletion
2
Hydrogel-Mediated Delivery of PD-1 Checkpoint Inhibitors
3
Synthetic Hydrogel Platform for Immune Checkpoint Inhibition
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All chemicals not otherwise specified were purchased from Sigma-Aldrich. For preparation of sterile MDP hydrogels, 2% wt/vol solutions were dissolved in 298 mmol/L sucrose to support cytocompatibility. Anti–PD-1 checkpoint inhibitor antibodies and IgG isotype controls were purchased from Bio X Cell. Peptide stock solutions (K2-MDP) were first prepared at 4% wt/vol in 298 mmol/L sucrose. The target final loading concentrations of checkpoint inhibitor antibodies within the gels were 300 μg anti–PD-1 per 60 μL gel (5 μg/μL for PD-1) or the same loading concentration of respective isotype IgG for negative control tests. The antibody stocks were provided in PBS, a buffer that is compatible for peptide hydrogelation via phosphate anion cross-linking of charged peptide nanofibers.
The volume of antibody stock solution (usually provided at stock concentrations of ∼7–9 μg/μL) required to achieve the final desired dose in the gel was then calculated, typically requiring a mixing of 1 part 4% wt/vol peptide with 2–3 parts antibody stock solution (with any leftover volume supplemented with additional PBS). The final antibody-loaded hydrogel formulations were 0.75X PBS, 75 mmol/L sucrose, 1% wt/vol peptide (10 mg/mL, ∼5–6 mmol/L), 5 μg/μL PD-1 or isotype control.
The volume of antibody stock solution (usually provided at stock concentrations of ∼7–9 μg/μL) required to achieve the final desired dose in the gel was then calculated, typically requiring a mixing of 1 part 4% wt/vol peptide with 2–3 parts antibody stock solution (with any leftover volume supplemented with additional PBS). The final antibody-loaded hydrogel formulations were 0.75X PBS, 75 mmol/L sucrose, 1% wt/vol peptide (10 mg/mL, ∼5–6 mmol/L), 5 μg/μL PD-1 or isotype control.
4
Establishing Esophageal Squamous Cell Carcinoma Xenografts
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To establish human esophageal squamous carcinoma cell xenografts, 2 × 106 ESCC cells suspended in PBS/Matrigel were injected subcutaneously into the right flank region of 6-week-old BALB/c nude mice. Tumor growth was monitored every 5 days until the subcutaneous tumors reached an average size of over 100 mm3. The mice were weighed every 5 days to monitor their general health. In pharmacological experiments, mice were intraperitoneally treated with either DY131 (40 or 80 mg/kg) or vehicle (1% DMSO + ddH2O) every 2 days for 25 days, starting on day 5 after tumor injection. For anti-programmed death 1 (PD1) treatments, mice were intraperitoneally administered mouse anti-PD1 antibodies (100 µg per mouse) or IgG isotype control (BioXCell) every 3 days for 25 days, beginning on day 5 after tumor injection. All animal procedures were conducted following the approved protocols of Fudan University’s Institutional Animal Care and Use Committee. Efforts were made to minimize the animals’ distress, pain, or discomfort throughout the study.
5
In Vivo Tumor Immunotherapy Evaluation
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Prior to treatments initiation, mice were randomly assigned to different groups with similar average tumor volumes. For in vivo treatments, VC (4 g/kg body weight; daily; i.p.) (Sigma) or PBS control, Sora (15 mg/kg body weight; daily; i.g.) (MCE); PD-L1 antibodies (75 μg per mouse; twice a week; i.p.) (BioXCell) or IgG isotype control, rIL-2 (1 μg per mouse; daily; i.p.) (R&D systems) or PBS control, and C-176 (750 nmol per mouse; daily; i.p.) (MCE) or solvent were injected for 2 weeks beginning on day 7 after the establishment of mice liver cancer models. To deplete CD8+ T cells in vivo, mice were intraperitoneally injected with 100 μg of anti-CD8 antibody (BioXCell) or IgG isotype control (BioXCell) 3 days and 1 day before tumor implantation and twice weekly thereafter to ensure sustained depletion of CD8+ T cell subset during the experimental period.
6
Comprehensive Cell Line Validation and Inhibitor Characterization
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All cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), independently validated by short tandem repeat (STR) DNA fingerprinting, and tested negative for mycoplasma infection. NCI-H157, A549, NCI-H1299, HeLa, BT549, MDA-MB-231, CT26, B16-F10, MC-38, LLC and 293T cells were cultured in RPMI 1640 or DMEM media, supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C in a humidified atmosphere with 5% CO2. Inhibitors for ATM (KU-60019, AZD1390, AZD0156), STING (H151), p-TBK1 (GSK8612), p-STAT1 (fludarabine) were purchased from Selleck (Houston, TX, USA). Human IFNβ and Galectin-9 ELISA assay Kits were purchased from R&D SYSTEMS (Minneapolis, MN, USA). Anti-human Gal-9 antibody was purchased from BioRad (Hercules, CA, USA). Anti-mouse Gal-9 antibody and IgG isotype control were purchased from BioXCell (Lebanon, NH, USA). Antibodies against cGAS, STING, p-STING(Ser366), p-TKB1(Ser172), TKB1, p-STAT1(Tyr701), STAT1 and Actin were purchased from Cell Signaling Technology (Cambridge, MA, USA). Anti-mouse fluorochrome-conjugated antibodies including FITC-CD4, Percp/Cy5.5-CD8, AF700-CD3, APC/Cy7-CD45, PE/Cy7-IFNγ antibodies were purchased from BioLegend (San Diego, CA, USA).
7
Investigating Radiation-Induced Tongue Damage
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Doses of anti-IL-1R (anti-Ly6G (BioXcell), and anti-G-CSF (R&D Systems) neutralizing antibodies were based on previous studies (31 (link)–33 (link)). Isotype control antibodies dose and administration schedules are included (Supplementary Table 2
8
Evaluation of Combination Therapies
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AZD1775, H151 and C176 were purchased from Selleck chemical (Houston, TX). Anti-PD-L1 antibodiy (clone 10F.9G2) and IgG isotype control were purchased from BioXcell (West Lebanon, NH). Cisplatin and etoposide were purchased from Accord Healthcare (Durham, NC). Recombinant IFN-γ was purchased from Biolegend (San Diego, CA).
9
Evaluating TOPK Inhibitors and Anti-PD-L1 in Renca Tumors
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Renca-WT and Renca-TOPK KD cells (1×106) were inoculated subcutaneously in the back of 6-8-week-old wild-type BALB/c mice. For the treatment group, Renca cell lines were inoculated with 1×106 cells subcutaneously on the back of 6-8-week-old wild-type BALB/c mice, and when the tumor volume grew to 50-100 mm3, the mice were randomly divided into four groups: control group, OTS964(TOPK inhibitors) treatment group, anti-PDL1 treatment group, and combination treatment group, respectively. InVivoMAb anti-mouse PD-L1 (Bio X Cell, West Lebanon, NH, USA) or IgG isotype control (Bio X Cell, West Lebanon, NH, USA) was administered to mice at a dose of 200ug/each by intraperitoneal injection every three days. For TOPK inhibitors, the tumors were administered orally at 40 mg/kg daily, while control mice received PBS orally. Tumor volumes were measured every three days along the long axis (a) and short axis (b), and tumor volumes were calculated using the following formula: V=ab2/2. Mice were sacrificed at the end of the experiment, tumors were excised and weighed, and tumor tissues were subsequently used for flow analysis, WB analysis, and immunohistochemistry, respectively.
10
Depletion of Neutrophils and Macrophages
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Neutrophil depletion was achieved with intraperitoneal injection of 150 μg anti-Ly6G Ab (BioXCell) or IgG isotype control (BioXCell) at day −1 and day 0 (day of infection). Macrophage depletion was performed with intravenous administration of 1 mg Clodronate liposomes (Clo-L) or 1 mg PBS liposomes (PBS-L) as control (Liposoma) at 1 day prior to infection.
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