The following antibodies were used in flow cytometry: phycoerythrin (PE)-conjugated anti-mouse CD44 (Biolegend, 103007), Sca-I (eBioscience, 12-5981-83), CD140a (Biolegend, 135906), CD13 (BD Pharmingen, 558745), MHC II (eBioscience, 12-5320-80), CD11b (eBioscience,12-0112-83), CD11c (eBioscience, 12-0114-82), CD86 (eBioscience, 12-0862-82), and CD45 (eBioscience, 12-0451-82). Antibodies used for western blotting analysis were: anti-p53 (FL-393; sc-6243, Santa Cruz Biotechnology); GAPDH (14C10, 2118, Cell Signaling Technology). Antibody used for Chip was: p53 (FL-393; sc-6243, Santa Cruz Biotechnology), IgG (normal mouse IgG, sc2025, Santa Cruz Biotechnology). The reagents for cell treatment were: Nutlin-3 (Nutlin-3, Selleck Chemicals, Houston, TX, USA), Cisplatin (Sigma-Aldrich), RANKL (recombinant mouse RANKL, 50 ng/ml, R&D), M-CSF (recombinant mouse M-CSF, 25 ng/ml, R&D), and OPG (recombinant OPG, 50 ng/ml, Sigma-Aldrich). RANKL for mouse injection (recombinant mouse RANKL, 2 mg/kg/day) was from R&D Systems.
Recombinant mouse rankl
Manufactured by R&D Systems
Sourced in United States
Recombinant mouse RANKL is a cytokine that plays a key role in the regulation of bone metabolism. It is a member of the tumor necrosis factor (TNF) ligand family and is essential for the formation, function, and survival of osteoclasts, the cells responsible for bone resorption.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant mouse m csf, by R&D Systems (15 mentions)
M csf, by R&D Systems (8 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Trap stain kit, by Merck Group (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Raw264.7 cell, by American Type Culture Collection (4 mentions)
Alpha minimal essential medium α mem, by Thermo Fisher Scientific (3 mentions)
Trap staining kit, by Merck Group (3 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (2 mentions)
Antibodies against nfatc1 and c fos, by Abcam (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
C fos, by Cell Signaling Technology (2 mentions)
Osteo assay surface for bone resorption, by Corning (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (2 mentions)
Primescript rt reagent kit, by Takara Bio (2 mentions)
Macrophage colony stimulating factor m csf, by R&D Systems (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Anti β actin, by Santa Cruz Biotechnology (2 mentions)
Nfatc1, by Abcam (2 mentions)
39 protocols using recombinant mouse rankl
1
Flow Cytometry and Western Blot Analysis of Cell Signaling Pathways
2
RANK Signaling in B Cell Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RANKK240E-expressing or wt B cells from littermates were stimulated with 100 ng/ml mouse recombinant RANKL (R&D Systems), 10 µg/ml neutralizing antibody (purified mouse α-RANKL; eBioscience), or 0.5 µM CpG (InvivoGen) for 1 to 48 h at 37°C. Following transduction, Bal17 cells were stimulated with 100 ng/ml mouse recombinant RANKL for 5 min to 48 h at 37°C. SP600125 was purchased from Sigma-Aldrich, and Cal-101 and PD0325901 (PD901) from Selleckchem, and BAY70-1182 was purchased from Caymanchem. The inhibitors were dissolved in DMSO and used at the indicated concentrations. Murine and human CLL cells were cultivated in coculture with murine bone marrow–derived stromal cells (ST-2) at a ratio of 10:1 and were treated with 10 µg/ml neutralizing antibody (purified mouse α-RANKL; eBioscience) for 24 to 72 h. To test RANKL dependency of transplanted RANKK240E-expressing CLL cells, peripheral blood cells derived from transplanted mice were stimulated with mouse recombinant RANKL (R&D Systems) and subsequently treated with neutralizing antibody (purified mouse α-RANKL; eBioscience) as described above and analyzed after 24 h and up to 10 d for survival via DAPI staining and flow cytometric analysis.
3
Osteoclastogenesis Assay in RAW264.7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW264.7 cells were purchased from the American Type Culture Collection (Rockville, MD). Fetal bovine serum (FBS) and Alpha minimal essential Medium (α‐MEM) were obtained from Gibco (life technologies, CA). Penicillin‐streptomycin solution was obtained from Hyclone (Thermo Scientific, MA). Recombinant Mouse M‐CS F and Recombinant Mouse RANKL were obtained from R&D Systems (Minneapolis, MN). Cell Counting Kit‐8 was obtained from Dojindo Molecular Technologies (Dojindo, Japan). TRAP stain kit was obtained from Sigma–Aldrich (St. Louis, MO). Actin Cytoskeleton and Focal Adhesion Staining Kit was purchased from Millipore (Darmstadt, Germany). Antibodies against P38, p‐P38, ERK, p‐ERK, JNK, p‐JNK, and β‐actin were purchased from Cell Signaling (Danvers, MA). ERK, JNK, and P38 MAP kinase inhibitors were purchased from Medchemexpress (Princeton, NJ). Bovine cortical bone slices for Pit formation assay were obtained from Boineslices.com (Jelling, Denmark). Antibodies against NFATC1 and c‐FOS were purchased from Abcam (Cambridge). Staphylococcal protein A was purchased from Sigma–Aldrich.
4
Osteoclast Differentiation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
α‐Minimum essential medium (α‐MEM) and foetal bovine serum (FBS) were obtained from Gibco; Thermo Fisher Scientific, Inc. Soluble recombinant mouse macrophage‐colony stimulating factor (M‐CSF) and recombinant mouse RANKL were obtained from R&D Systems, Inc. Tartrate‐resistant acid phosphatase (TRAP) was obtained from Sigma‐Aldrich and Merck KGaA. High purity Ti particles (average diameter. 1‐5 µm) were obtained from Johnson Matthey (cat. no., 00681). The antibodies (GAPDH, NFkB, C‐FOS, NFATc1, p‐p38,) were purchased from Cell Signaling Technology, Inc. Enzyme‐linked immunosorbent assay (ELISA) kits (IL‐6, IL‐1β, TNF‐α, IL‐10, Arg‐1, iNOS) were purchased from R&D Systems, Inc Flow cytometry anti‐mouse CD16/32‐PE (cat. no.553145) and anti‐mouse CD206‐Alexa 647 (cat. no.565250) were purchased from BioLegend Inc The p38α/β MAPK inhibitor (SB202190) was purchased from Selleck.
5
Osteoclastogenesis Signaling Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant mouse M-CSF and recombinant mouse RANKL were purchased from R&D Systems, USA. Primary antibodies to c-Jun, Akt, MAPK p38, ERK 1/2, phospho-ERK 1/2, IKK β, and phospho-IKK β were obtained from Cell Signaling Technology (CST), Danvers, MA. Primary antibody to β-actin, anti-mouse, and anti-rabbit HRP conjugated secondary antibodies were purchased from Sigma-Aldrich, USA. Female Swiss albino mice (18–22 g) were procured and maintained at the animal house facility of ACTREC. All in vivo protocols and experiments were carried out in accordance with the relevant guidelines approved by the Institutional Animal Ethics Committee, ACTREC. Animals were fed with standard laboratory diet and water ad libitum.
6
Osteoclast Differentiation and Bone Resorption
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW264.7 cells were obtained from the American Type Culture Collection (Rockville, MD, USA). Recombinant mouse RANKL and recombinant mouse M-CSF were purchased from R&D Systems (Minneapolis, MN, USA). The Osteo Assay Surface for Bone Resorption was purchased from Corning (NY, USA). The TRAP stain kit was obtained from Sigma-Aldrich (NY, USA). The Actin Cytoskeleton and Focal Adhesion Staining Kit was purchased from Millipore (Darmstadt, Germany). Alpha minimal essential medium (α-MEM) and fetal bovine serum (FBS) were purchased from Gibco (Life Technologies, USA). Penicillin-streptomycin solution was obtained from Hyclone (Thermo Scientific, USA). Membrane dye DiI and Cell Tracker Green were obtained from Life Technologies.
7
VSMC Osteogenic Differentiation Assay
8
Tectorigenin Inhibits Osteoclastogenesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tectorigenin (purity >98% by HPLC) was obtained from Tauto Biotech (Shanghai, China). Recombinant mouse RANKL and M‐CSF were purchased from R&D Systems (Minneapolis, MN, USA). Alpha Modification of Eagle's Medium (α‐MEM), foetal bovine serum (FBS), penicillin, and streptomycin were purchased from Gibco (Rockville, MD, USA). Cell Counting Kit‐8 (CCK‐8) was purchased from Dojindo Molecular Technology (Japan). Tartrate‐resistant acid phosphatase (TRAP) staining kit, dimethyl sulphoxide (DMSO), ethylenediaminetetraacetic acid (EDTA) were obtained from Sigma‐Aldrich (St Louis, MO, USA). NF‐κB p65 (CST 8242), phosphor‐NF‐κB p65 (CST 3033), IκBα (CST 4812), and phosphor‐IκBα (CST 2859) antibodies were purchased from Cell Signalling Technology (Beverly, MA, USA). β‐actin (SC‐47778) antibodies was purchased from Santa Cruz Biotechnology.
9
Osteoclastogenesis Inhibition by DHA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant Mouse RANKL and Recombinant Mouse M-CSF were purchased from R&D Systems (Minneapolis, MN, USA). Antibodies against AIF, cytochrome c, cleaved caspase-3, Bax, Bcl-2 and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Osteo Assay Surface for Bone Resorption was purchased from Corning (Corning, NY, USA). Bovine cortical bone slices were obtained from Boineslices.com (Jelling, Denmark). Cell Counting Kit-8 was obtained from Dojindo Molecular Technologies (Dojindo, Japan). The TRAP stain kit was obtained from Sigma-Aldrich (St. Louis, MO, USA). The Actin Cytoskeleton and Focal Adhesion Staining Kit was purchased from Millipore (Darmstadt, Germany). The DCFDA cellular ROS detection assay kit was obtained from ABcam (Cambridge, UK). Alpha minimal essential Medium (α-MEM) and fetal bovine serum (FBS) were purchased from Gibco (Life Technologies, Carlsbad, CA, USA). Penicillin–streptomycin solution was obtained from Hyclone (Thermo Scientific, Waltham, MA, USA). DHA was purchased from Sigma-Aldrich.
10
Mouse Macrophage Activation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All chemical reagents were purchased from Sigma-Aldrich (USA). The peptides were purchased from China Peptides Co. Ltd. (purity: 95%). All the primary antibodies were purchased from Cell Signaling Technology Inc., (USA) and the secondary antibody was bought from Beyotime Biotechnology (China). The blocking buffer and primary antibody dilution buffer for western blot were obtained from Beyotime Biotechnology. All cell culture reagents were purchased from Thermo Fisher Scientific Inc. (GibcoTM, USA). The mouse macrophage RAW 264.7 cell was obtained from the American Type Culture Collection (ATCC). Recombinant Mouse RANKL was purchased from R&D systems, Inc. (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!