To explore the role of PD-1 and PD-1 ligands in ISO-induced stress-induced cardiomyopathy, anti-PD-1, anti-PD-L1, anti-PD-L2, or isotype control Abs were administered IP at a dose of 300 μg per mouse.11 (link),12 (link) The following Abs (Bio X Cell) were employed: rat anti-PD-1 (clone RMP1-14); rat anti-PD-L1 (clone 10F.9G2); rat IgG2b (clone LTF-2), isotype control for anti-PD-L1; rat anti-PD-L2 (clone TY25); and rat IgG2a (clone 2A3), isotype control for anti-PD-1 and anti-PD-L2.11 (link) All Abs were diluted in InVivoPure dilution buffers (Bio X Cell) to a concentration of 1 mg/mL just prior to use, according to the manufacturer’s instructions. Mice received Ab injections 2 days before, and 1 and 4 days after ISO administration.
Invivopure dilution buffer
Manufactured by BioXCell
Sourced in United States
InVivoPure dilution buffer is a sterile, ready-to-use solution designed to dilute samples for in vivo applications. It is formulated to maintain the stability and viability of biological samples during preparation and storage.
Automatically generated - may contain errors
Lab products found in correlation
Yts 169, by BioXCell (1 mentions)
Clone ltf 2, by BioXCell (1 mentions)
Clone gk1, by BioXCell (1 mentions)
Matrigel, by Corning (1 mentions)
Fulvestrant, by Bio-Techne (1 mentions)
Dacomitinib, by Selleck Chemicals (1 mentions)
Mouse tnf α, by R&D Systems (1 mentions)
Mouse vegf elisa kit, by R&D Systems (1 mentions)
Rat igg2a clone 2a3, by BioXCell (1 mentions)
Recombinant mouse il 22, by Thermo Fisher Scientific (1 mentions)
6 protocols using invivopure dilution buffer
1
Role of PD-1/PD-L Axis in ISO-Induced Cardiomyopathy
2
Preventing T Cell Activation in Stress-Induced Cardiomyopathy
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To prevent T cell activation in ISO-induced stress-induced cardiomyopathy, recombinant cytotoxic T lymphocyte-associated antigen-4 (CTLA4) Ig or recombinant human isotype control IgG1 Ab were administered IP, at a dose of 200 μg per mouse.13 (link),14 (link) The Abs were obtained from Bio X Cell and diluted in InVivoPure dilution buffers (Bio X Cell) to a concentration of 1 mg/mL just prior to use, according to the manufacturer’s instructions. Mice received injections 2 days before and 1 and 4 days after ISO administration.
3
Breast Cancer Metastasis Mouse Model
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Mouse experiments were performed according to Good Scientific Practice-principles and approved by the Ethical Committee (EC) of the Faculty of Veterinary Medicine, Ghent University (EC 2020-030 and 2021-018).
Eight-week (w)-old female and male BALB/c mice were mated and pups were weaned 12-14 days (d) post-parturition. One hour (h) after weaning, lactating females were intraductally inoculated in the third mammary gland pair with 5 × 104 4T1-luc cells suspended in a 100 μl mixture of PBS and Matrigel® (1:10; Corning, Bedford, MA, USA) under inhalation anesthesia and analgesia as described previously (15 (link)). For intravenous (i.v.) treatment, eMSCs or eEpSCs were dissolved in DMEM at 3x105 per 100 μl and injected through the tail vein using a 29G insulin needle. I.v. injections with DMEM only were used as a negative (sham) control. Antibody treatments for depletion of CD4+ (clone GK1.5) and CD8α+ T-cells (clone YTS169.4) or rat IgG2b isotype controls (clone LTF-2) were purchased from BioXCell (West Lebanon, NH, USA) and diluted (200 μg/100 μl) in InVivoPure dilution buffer (BioXCell) at pH6.5 (for anti-CD4) or pH7 (for anti-CD8α and isotype control) for i.p. administration.
Eight-week (w)-old female and male BALB/c mice were mated and pups were weaned 12-14 days (d) post-parturition. One hour (h) after weaning, lactating females were intraductally inoculated in the third mammary gland pair with 5 × 104 4T1-luc cells suspended in a 100 μl mixture of PBS and Matrigel® (1:10; Corning, Bedford, MA, USA) under inhalation anesthesia and analgesia as described previously (15 (link)). For intravenous (i.v.) treatment, eMSCs or eEpSCs were dissolved in DMEM at 3x105 per 100 μl and injected through the tail vein using a 29G insulin needle. I.v. injections with DMEM only were used as a negative (sham) control. Antibody treatments for depletion of CD4+ (clone GK1.5) and CD8α+ T-cells (clone YTS169.4) or rat IgG2b isotype controls (clone LTF-2) were purchased from BioXCell (West Lebanon, NH, USA) and diluted (200 μg/100 μl) in InVivoPure dilution buffer (BioXCell) at pH6.5 (for anti-CD4) or pH7 (for anti-CD8α and isotype control) for i.p. administration.
4
Examining TGFβ Inhibition in Col4a1 Mutant
5
Combination Therapy Protocol for Cancer
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Dacomitinib (Selleckchem, Houston, TX, USA) and fulvestrant (Tocris, Minneapolis, MN, USA) were dissolved in DMSO. DMSO, as a control, was given at the same amount as the maximum DMSO used in treatment groups. For in vivo studies, peanut oil was used to dilute the dose of fulvestrant for s.c. administration. Dacomitinib was further dissolved in polyethylene glycol (30%) and PBS for oral gavage administration. Rat monoclonal blocking anti-mouse PD-1 (clone RMP1.14) and isotype control (rat IgG2a, clone 2A3) were purchased from BioXcell and prepared in InvivoPure dilution buffer (BioXcell, Lebanon, NH, USA). Mouse IFN-ϒ, mouse TNF-α, and mouse VEGF ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA).
6
Therapeutic Supplementation of IL-22 in Bone Repair
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Mice were osteotomized as described above, and treatment was started 24 h post-surgery to mimic the feasibility of this treatment approach under clinical conditions. The supplementation and neutralization of IL-22 were performed via intraperitoneal injection of recombinant mouse IL-22 (PeproTech, Rocky Hill, NJ, USA) or rat anti-mouse functional grade IL-22 monoclonal antibody (clone IL22JOP, Thermo Fisher, Waltham, MA, USA). IL-22 supplementation was performed with 5 µg protein in 100 µl InVivoPure dilution buffer (BioXCell, Lebanon, NH, USA) every second day, with the last injection timepoint 1 week post-surgery. The neutralizing IL-22 antibody was diluted to 100 µg in 100 µl InVivoPure dilution buffer, with a treatment schedule similar to that described for IL-22 supplementation.
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