PMMA particles with mean diameter 6 μm (range 0.1–10 μm) were purchased from a commercial resource (Polysciences, Warrington, PA). Endotoxin was removed from the particles according to the protocol described by Ragab et al. [15 (link)]. The particles were rinsed in ethanol four times and sterilized in 70% ethanol with shaking overnight. The particles were then rinsed four times with PBS and resuspended in serum-free α-MEM. The removal of endotoxin was confirmed by a Limulus amoebocyte lysate assay (Cat number QCL-1000; BioWhittaker, Walkersville, MD) with sensitivity of 0.005 endotoxin unit (EU)/mL.
Pmma particles
Manufactured by Polysciences
PMMA particles are a type of lab equipment used for various applications in scientific research and industrial processes. These particles are made of polymethyl methacrylate (PMMA), a transparent and durable plastic material. PMMA particles are available in different sizes and surface characteristics to suit different requirements. The core function of PMMA particles is to serve as a versatile material for various experimental and analytical purposes.
Automatically generated - may contain errors
Lab products found in correlation
Limulus amoebocyte lysate assay, by Lonza (1 mentions)
Limulus amebocyte lysate kit, by Lonza (1 mentions)
Recombinant human m csf, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Miltenyi Biotec (1 mentions)
Msu crystal, by InvivoGen (1 mentions)
Rankl, by Thermo Fisher Scientific (1 mentions)
Lymphoprep, by STEMCELL (1 mentions)
Recombinant human il 6, by Miltenyi Biotec (1 mentions)
4 protocols using pmma particles
1
Preparation of Endotoxin-Free PMMA Particles
2
PMMA Particle Preparation and Curcumin Dissolution
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Commercially available PMMA particles (Polysciences) with a mean diameter of 4.8 μm (0.1–16) were used in this study as described previously [19 (link)]. Ninety percent of the particles were <10 μm in diameter. Adherent endotoxin in particles was removed by sterilization in 70% ethanol for 48 h and washing with sterile phosphate-buffered saline (PBS) thrice. The absence of endotoxin in PMMA particles was confirmed using a Limulus Amebocyte Lysate Kit (BioWhittaker, Walkersville, MD) at a detection level of <0.05 EU/ml. The PMMA particles were then suspended in sterile PBS at a concentration of 1 mg/ml. CUR with ≥98% purity was obtained from Aladdin Industrial Incorporation (Ontario, CA, USA). A total of 1 mg CUR was dissolved in 0.5 ml dimethyl sulfoxide (DMSO) at a concentration of 2 mg/ml and stored at −20°C until needed.
3
Sterilization and Preparation of PMMA Particles
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PMMA particles (Polyscience) were sterilized before injection by washing with 70% EtOH for three times followed by 3 times in PBS and finally resuspended in PBS (0.2 mg/ml for in vitro studies and 20 ul per injection of 5 mg/ml in vivo). All FACS antibodies, buffers and reagents were purchased from either BD Biosciences, eBioScience/Thermo Fisher or BioLegend.
4
Osteoclastogenesis Inhibition Protocol
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The Syk inhibitor (R788) was obtained from AdooQ BioScience (Irvine, CA), p38 (SB203580), JNK (SP600125), MEK/ERK (PD98059) inhibitors and MSU crystals were from Invivogen (San Diego, CA).
PMMA particles (diameter range 1e10 mm) were obtained from Polysciences, Inc. (Warrington, PA). BCP crystals in the form of hydroxyapatite (HA) were synthesized by alkaline hydrolysis of brushite as described 38 . Recombinant human IL-6 and IFN-g were obtained from Miltenyi Biotech (Bergisch Gladbach, Germany). Recombinant human M-CSF and RANKL were from PeproTech (Rocky Hill, NJ). Lymphoprep was obtained from Stemcell Technologies (Grenoble, France). All primary antibodies were obtained from Cell Signaling Technology (Beverly, Massachusetts). Secondary antibodies, cell culture reagents, Acid Phosphatase, Leukocyte (TRAP) Kit and all chemicals were from SigmaeAldrich (St. Louis, Missouri).
PMMA particles (diameter range 1e10 mm) were obtained from Polysciences, Inc. (Warrington, PA). BCP crystals in the form of hydroxyapatite (HA) were synthesized by alkaline hydrolysis of brushite as described 38 . Recombinant human IL-6 and IFN-g were obtained from Miltenyi Biotech (Bergisch Gladbach, Germany). Recombinant human M-CSF and RANKL were from PeproTech (Rocky Hill, NJ). Lymphoprep was obtained from Stemcell Technologies (Grenoble, France). All primary antibodies were obtained from Cell Signaling Technology (Beverly, Massachusetts). Secondary antibodies, cell culture reagents, Acid Phosphatase, Leukocyte (TRAP) Kit and all chemicals were from SigmaeAldrich (St. Louis, Missouri).
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