Ag3PO4 microparticles (S1–S4) were prepared through a sonochemical and hydrothermal method. In a typical synthesis procedure, AgNO3 (S1: 1.25, S2: 7.5, S3: 12.5, and S4: 25.0 mmol) was dissolved in 45 mL of deionized water. The prepared Na3PO4 solution (2.50 mmol, 5 mL) was then added to the preceding solution under ultrasonic irradiation at room temperature. After 30 min of ultrasonic irradiation (Misonix, Inc., Farmingdale, NY, USA: XL-2020, 20 kHz, 600 W), the mixture was transferred into a Teflon-lined stainless steel autoclave. The autoclave was sealed and heated in an electric oven at 180 °C for 24 h. After the autoclave naturally cooled to room temperature, the precipitates were centrifuged and washed three times with ethanol and deionized water, and then dried in vacuum at 50 °C for 8 h.
Xl2020
Manufactured by Bioventus
Sourced in United States
The XL2020 is a versatile lab equipment designed for various scientific applications. It features advanced technology and precise measurements to support laboratory workflows. The core function of the XL2020 is to provide accurate and reliable data, enabling researchers to conduct their experiments with confidence.
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Lab products found in correlation
Optison, by GE Healthcare (3 mentions)
Tecnai g2 spirit twin, by Thermo Fisher Scientific (3 mentions)
Multisizer 3, by Beckman Coulter (2 mentions)
Cd sprague dawley igs rat, by Charles River Laboratories (1 mentions)
End it dna end repair kit, by Illumina (1 mentions)
Hiseq platform, by Illumina (1 mentions)
Formaldehyde, by Merck Group (1 mentions)
Polyethylene glycol stearate, by Merck Group (1 mentions)
Multisizer 2, by Beckman Coulter (1 mentions)
1 2 distearoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
1 2 distearoyl sn glycero 3 phosphoethanolamine n biotinyl polyethylene glycol 2000, by Avanti Polar Lipids (1 mentions)
D pbs, by Nacalai Tesque (1 mentions)
24 protocols using xl2020
1
Synthesis of Silver Phosphate Microparticles
2
Microbubble Formulation and Characterization
3
Culturing and Treating Cortical Neurons
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P1 mouse or rat cortical neurons were isolated and cultured essentially according to a previously published protocol [51 (link)]. At 7 d in vitro (DIV), neurons were treated with or without 140 nM αSyn monomer or PFF. Treated neurons were cultured as usual and collected after 8–9 d or after a specified time. PFF was sonicated before each usage with a water-bath cup-horn sonicator (Misonix XL2020) at 50% power for 5 min. For proteasomal inhibition, primary neurons were treated with DMSO or 100 nM epoxomicin for 8 h, and after incubation, cell lysates or homogenates were prepared.
4
Intramuscular PFF Injection in Mice
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C57BL/6J mice were from Van Andel Institute internal colony. CD (Sprague Dawley) IGS rats were obtained from Charles River. Transgenic mice expressing human A53T αSyn from the mouse prion protein promoter (line G2–3, kindly provided by Dr. Michael Lee, University of Minnesota) were described previously [24 (link)]. The bilateral intramuscular PFF injection was performed according to a previously published protocol [43 (link)] with slight modification. Briefly, the PFF used for the intramuscular injection was prepared by shaking 350 μM αSyn at 1000 rpm for 2 d at 37 °C. PFF was aliquoted and stored at − 80 °C. Before each injection, PFF was diluted to 0.1 μg/μL and sonicated with the water-bath cup-horn sonicator (Misonix XL2020) at 25% power for 2 h, and then 10 μL of PFF (0.1 μg/μl) or 10 μL PBS was injected into biceps femoris on each hindlimb.
5
Intracerebral Injection of α-Synuclein PFFs in Mice
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Prior to injection, human α-syn PFFs were thawed and then sonicated at RT with a water-bath cup-horn sonicator (Misonix XL2020, 50% power, 120 pulses 1 s ON, 1 s OFF). Post-sonication fibril morphology was validated by TEM imaging protocol as described above. Prior to injection, 4-week-old mice (both sexes) were anesthetized with an isoflurane/oxygen mixture and injected unilaterally with either 2μL of PFFs (5μg/μl) or 2μL of PBS as a control in the dorsal striatum (coordinates from bregma: AP: +0.2 mm; ML: – 2.0 mm; DV: – 2.6 mm from dura) at a rate of 0.2μL/min using a glass capillary attached to a 10μL Hamilton syringe. After injection, the capillary was left in place for 4 minutes before being slowly removed.
6
Formaldehyde Crosslinking and Input-Seq
7
Albumin Microbubble Preparation
8
In vitro Aggregation and Application of Alpha-Synuclein Fibrils
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PFFs or α-syn monomer treatments were performed after the brain slices were cultured in vitro for 5 days. Recombinant mouse or human α-syn was purified following the protocol reported previously [43, 44 (link)]. To prepare amyloid fibrils, purified α-syn was dialyzed against phosphate-buffered saline (PBS), concentrated to 350 μM and shaken at 37°C, 1,000 rpm for 7 days. PFFs were diluted to 7 μM in sterilized PBS and sonicated at 50% power for 5 minutes (Misonix XL2020). The morphology of PFFs was imaged by a Tecnai G2 Spirit TWIN transmission electron microscopy (Supplementary Figure 2 ). Immediately after sonication, 7 μM (0.1 μg/ μl) PFFs were added onto the slices (15 μl/a half rat cerebellar slice; 5 μl/a half mouse cerebellar slice; 40 μl/a rat coronal cortical slice). For α-syn monomer treatment, the same amount of soluble α-syn monomer was added onto the slices. After 2 days, PFFs/PBS solution was aspirated and half of the culture medium was replaced with fresh medium. The slices were collected 2, 3, or 4 weeks after PFFs or monomer treatment as indicated.
9
Neuronal Fractionation and Extraction
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At DIV 16, neurons were homogenized into TBS buffer (50 mM Tris, 150 mM NaCl, pH 7.5), sonicated at 50% power for 5 min (Misonix XL2020), and centrifuged at 100,000×g for 30 min at 4 °C. The pellets were resuspended in 1% Triton-100 in TBS, sonicated, and centrifuged as described above. The supernatant was collected as the Triton-soluble fraction. The pellet was resuspended in 2% SDS in TBS.
10
Perfluorobutane Microbubbles for Sonothrombolysis
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