Zoledronate, galangin, icariside II, kaempferol, quercetin, Zoledronate, galangin, icariside II, kaempferol, quercetin (Figure 1 ), and 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyl tetrazolium bromide reagents were purchased from Sigma (St. Louis, MO), and 3‐methyladenine (3‐MA; a type‐III phosphatidylinositol 3‐kinase [PI‐3 K] inhibitor) and rapamycin reagents were obtained from Calbiochem (La Jolla, CA). The antibodies against beclin‐1 and Autophagy protein 5 (ATG5) were purchased from Cell Signaling Technology (Beverly, MA). The antibodies against collagen I, osteocalcin, bone morphogenetic protein 2 (BMP‐2), Osterix, and RUNX2 were purchased from ABCam (Cambridge, MA). The antibodies against LC3 were purchased from Sigma. SQSTM1/p62, β‐actin antibody, mouse anti‐rabbit IgG antibody, and rabbit anti‐mouse IgG antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). All other chemicals and reagents were purchased from Sigma unless otherwise specified.
Zoledronate
Manufactured by Merck Group
Sourced in United States, Australia
Zoledronate is a laboratory reagent that is used as a standard in various analytical procedures. It is a crystalline solid that is soluble in water and commonly used as a reference standard to quantify and identify related substances in pharmaceutical and biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Alendronate, by Merck Group (5 mentions)
Clodronate, by Merck Group (3 mentions)
Pravastatin, by Merck Group (3 mentions)
Ibandronate, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Mouse anti rabbit igg antibody, by Santa Cruz Biotechnology (2 mentions)
Human ab serum, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Prism 7, by GraphPad (1 mentions)
Lymphoprep, by STEMCELL (1 mentions)
Interleukin 2 (il 2), by BioLegend (1 mentions)
Tracp, by Merck Group (1 mentions)
M csf, by Thermo Fisher Scientific (1 mentions)
Easysep human gamma delta t cell isolation kit, by STEMCELL (1 mentions)
Recombinant human interleukin 2, by Thermo Fisher Scientific (1 mentions)
Ficoll paque, by GE Healthcare (1 mentions)
Ldh cytotoxic assay kit, by Thermo Fisher Scientific (1 mentions)
96 well plate, by Thermo Fisher Scientific (1 mentions)
Alamarblue assay, by Thermo Fisher Scientific (1 mentions)
27 protocols using zoledronate
1
Autophagy Regulation in Osteogenesis
2
PBMC expansion with zoledronate and IL-2/IL-15
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMC were resuspended in Roswell Park Memorial Institute (RPMI) supplemented with 10 % heat-inactivated human AB serum (Invitrogen, Merelbeke, Belgium), zoledronate (5 μM; Sigma-Aldrich, Diegem, Belgium), IL-2 (100 IU/mL), and/or IL-15 (100 IU/mL) at a final concentration of 1 × 106 cells/mL. Cell cultures were maintained at a cell density of 0.5–2 × 106 cells/mL and were replenished every 2 to 3 days by adding IL-2/IL-15-supplemented medium. Phenotypic and functional assays were performed on cells harvested at least 14 days after first stimulations.
3
Isolation and Expansion of Vδ2+ T Cells
4
In Vitro Osteoclastogenesis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole bone marrow was flushed from the tibia of 6-8 week old male or female C57BL/6 recombinase activating gene-2 null (Rag2−/−) mice, and cultured in the presence of α-MEM and 25ng/ml macrophage colony stimulating factor (M-CSF#315-02, Peprotech) for 3 days. Non-adherent bone marrow cells were maintained in osteoclastogenic medium (100 ng/ml RANKL #47187000, OYC Americas and 20 ng/ml M-CSF) for 7 days. BMMPIs or Zoledronate (#SML0223, Sigma Aldrich) were added at varying concentrations (1 to 50μM) for 24 hours. Vehicle control wells were treated with 0.5% DMSO that reflected the final concentration of the vehicle in the 50μM drug treated groups. Subsequently, osteoclasts were fixed with 4% paraformaldehyde, and TRAcP stained (387A, Sigma-Aldrich) as previously described [48 (link)]. Multinucleated osteoclasts were counted using ImageJ.
5
Vγ9Vδ2 T Cell Isolation and Expansion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs were isolated from the healthy volunteers’ whole blood (The First Affiliated Hospital of Jinan University), following standard Ficoll−Paque-based (GE Healthcare) density gradient centrifugation protocol. Vγ9Vδ2 T cells were generated with Zoledronate (Sigma-Aldrich) as described (40 (link)). Briefly, isolated PBMCs were suspended in RPMI 1640 medium supplemented with 10% FBS, 100 U ml−1 recombinant human interleukin-2 (Peprotech), and Zoledronate (50 μM). The PBMCs were seeded into 24 well plates, followed by routine culture procedures for 10 days before further tests. Vγ9Vδ2 T cells were maintained at a cell density of 1×106 ml−1. When ratio of Vγ9Vδ2 T cells out of total CD3+ cells reached 90%, they could be used in experiments. Vγ9Vδ2 T cells were characterized with PerCP-conjugated antihuman TCR Vδ2 (BioLegend) and V500-conjugated antihuman CD3 (BD Biosciences) via flow cytometry. In some experiments, the Vγ9Vδ2 T cells were further purified by negative selection with EasySep™ Human Gamma/Delta T Cell Isolation Kit (STEM CELL), and the purity of enriched Vγ9Vδ2 T cells was validated by flow cytometry and was generally higher than 98%.
6
Cytotoxicity of γδ T Cells Against Tumor Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytotoxic potential of γδ T cells against panel of tumor cell lines, oral tumor cell line (AW13516), colon tumor cell line (COLO-205), and B lymphoblastic cell line (Raji) was performed using lactate dehydrogenase (LDH) release assay as described previously (37 (link)). Tumor cell lines were treated for 18 h with zoledronate (100 µM; Sigma-Aldrich). γδ T cells were treated with HDMAPP (1 nM) and rIL-2 (50 IU/ml) in presence and absence of HDAC inhibitors, VPA (2 mM), TSA (100 nM), and SAHA (1 µM) for 72 h at 37°C were used as effectors. Additionally, for PD-1 blockade, anti-PD1 antibody (3 µg/ml) was added to HDAC inhibitor treated γδ T cells for 72 h at 37°C and were also used as effectors. Briefly, tumor cell lines were cocultured with effectors at 40:1 effector target (E/T) ratio for 4 h at 37°C in 96-well plates (Nunc, Denmark). After 4 h of coculture, an aliquot of 50 µl of media was used in LDH cytotoxic assay using the LDH cytotoxic assay kit (Thermo Fisher Scientific, USA) according to manufactures protocol. γδ T cell cytotoxicity was defined as % specific lysis = Experimental value – Effector cells spontaneous control − Target cells spontaneous control/Target Cell Maximum Control − Target cells spontaneous control.
7
Bisphosphonate Effects on pMSC Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
pMSCs (100 μl @ 2 × 104 cells/ml) were plated onto a 96 well microtitre-plate and allowed to attach for 24 hrs. The cells were subsequently incubated in 100 μl aliquots of endothelial growth media (EGM-2, Lonza, Mount Waverley VIC, Australia) with 0.25–200 μM clodronate, alendronate and zoledronate (Sigma-Aldrich, castle Hill, NSW, Australia) for 10 days. The proportion of viable cells and the proliferation rate compared to untreated (control) cells was determined by alamarBlue® assay (Life Technologies, Mulgrave, VIC, Australia).
8
Modulation of PBMC Immune Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1ml of PBMCs (1x106/ml) from three healthy donors were incubated for 10 days with either 2x1010E. coli MG1655 Δpal ΔlpxM OMVs (20000:1 ratio), 5 μM zoledronate (zoledronic acid monohydrate, Sigma-Aldrich) or PBS. In all cases, cells were also supplemented with 100 IU/ml IL-2 (human IL-2 IS premium grade, Miltenyi Biotec), which continued every three days along with re-adjustment of media. At day 3, 500 μl media was added to each well to increase the usable volume. Upon completion of each incubation, PBMCs in each sample were stained for various markers and analysed by flow cytometry (Supplementary Table 1
9
Synergistic Effects of Zoledronate and Pravastatin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The combination (ZOPRA) of an anti-osteoporosis bisphosphonate (zoledronate) and an anti-cholesterolemic statin (pravastatin) has been shown to accelerate the RIANS [39 (link)]. The ZOPRA treatment was applied under conditions that are described elsewhere. Briefly, cells were incubated with 1 µM pravastatin (#P4498, Sigma-Aldrich France, Saint-Quentin-Fallavier, France) in phosphate-buffered saline (PBS) (#14040-091, Sigma-Aldrich) for 24 h at 37 °C. Thereafter, 1 µM zoledronate (#SML0223, Sigma-Aldrich) in PBS was added to the culture medium, and cells were incubated for 12 h at 37 °C [39 (link)].
10
Osteoclast Differentiation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Clodronate was purchased from Cayman (E. Ellsworth, MI, USA) and dissolved in phosphate buffer saline (PBS). Zoledronate was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in phosphate buffer saline (PBS). AG490 was purchased from Calbiochem (La Jolla, CA, USA) and dissolved in dimethyl sulfoxide (DMSO). Control-IgG (Rat IgG2a Control) and IL-6-IgG (Anti-mIL-6-IgG) were purchased from InvivoGen (San Diego, CA, USA). Antibodies against phosphor-STAT3 and STAT3 were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-RANKL antibody was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The Leukocyte Acid Phosphatase (TRAP) kit, Leukocyte Alkaline Phosphatase (ALP) kit, and anti-β-actin antibody were obtained from Sigma-Aldrich (St. Louis, MO, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!