Phe0023

Manufactured by Thermo Fisher Scientific

The PHE0023 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It serves as a general-purpose device for various laboratory applications. The core function of this equipment is to facilitate specific tasks within the laboratory environment. No further details or interpretations are provided.

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Lab products found in correlation

Neo scmos camera, by Oxford Instruments (2 mentions) Cfi plan fluor 40 na 0.6 air objective, by Oxford Instruments (2 mentions) Alexa 488 conjugated goat anti mouse antibody, by Thermo Fisher Scientific (1 mentions) A32723, by Thermo Fisher Scientific (1 mentions) Tritc conjugated goat anti rabbit antibody, by Jackson ImmunoResearch (1 mentions) Alexa 647 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Nis element ar analysis software, by Nikon (1 mentions) 35 mm glass bottom dishes, by MatTek (1 mentions) Cellsens dimension system, by Olympus (1 mentions) Uplanapo 100 1.5 oil objective, by Olympus (1 mentions) Csu w1 confocal scanner, by Yokogawa (1 mentions) Eclipse ti, by Nikon (1 mentions) Sylgard 184, by Dow (1 mentions) Dynabeads m 450, by Thermo Fisher Scientific (1 mentions)

5 protocols using phe0023

CD44 and FGFR2 Colocalization in HUVECs

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HUVECs were plated on fibronectin (10 µg/mL, Thermo Fisher, PHE0023) -coated coverglass in a 48-well cell culture plate. Cells were washed with PBS, fixed with 3% paraformaldehyde, and permeabilized with 0.1% Brij 98. Next, the cells were blocked using 10% goat serum and incubated with CD44 rabbit antibody (Proteintech, 15675-1-AP, 1:200) and FGFR2 mouse antibody (Huabio, M1501-2, 1:50) at 4 °C overnight, followed by Alexa 488-conjugated goat anti-mouse antibody (Invitrogen, A32723, 1:500) and Alexa 647-conjugated goat anti-rabbit antibody (Invitrogen, A32733, 1:500) incubation for 1 h. The cell nuclei were stained with DAPI.
Primary MVECs were cocultured with 10 µg/mL plasma exosomes within 0 min, 15 min, and 30 min. Then, the cells were fixed, permeabilized, and blocked as described above. Next, the cells were incubated with EEA1 mouse antibody (CST, 48,453, 1:100) and CAV1 rabbit antibody (Abcam, ab32577, 1:250), followed by Alexa 647-conjugated goat anti-mouse antibody (Invitrogen, A32723, 1:500) and TRITC-conjugated goat anti-rabbit antibody (Jackson ImmunoResearch, 111-025-003, 1:100) incubation. Colocalization was analyzed by Nikon NIS-Element AR Analysis software. Pearson’s correlation was used to quantify the degree of colocalization between fluorophores.

Zhang Q., Chen L., Huang L., Cheng H., Wang L., Xu L., Hu D., He C., Fu C, & Wei Q. (2022). CD44 promotes angiogenesis in myocardial infarction through regulating plasma exosome uptake and further enhancing FGFR2 signaling transduction. Molecular Medicine, 28, 145.

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Evaluating CHD1L Silencing on GC Invasion

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Single-cell suspensions (100 µL) of BGC-823 cells infected with CHD1L-shRNA-1 or shRNA-NC or untreated cells (5×10⁵ cells/mL in serum-free medium) and 500 µL fibronectin (PHE0023; 80 ng/mL; Thermo Fisher Scientific, Inc.) were added to the upper and lower Matrigel-coated Transwell chambers, respectively, and cultured at 37 °C with 5% CO₂ for 24 h. After removing the Matrigel and non-invasive cells from the membrane, cells were fixed in precooled methanol for 30 min at 4 °C, followed by 5 min of Giemsa staining (1 mg/mL) at room temperature, ethanol dehydration (80%, 95%, and 97%), and gradient elution with a 20 min incubation at room temperature for each step. The number of invasive cells in five randomly selected high-power microscopic fields (magnification, ×200) was counted under an inverted microscope. The inhibitory rate of CHD1L silencing in GC cells was calculated as follows: inhibitory rate (%) = (control − sample)/control ×100.

Li D., Li C., Wang Y., Wang Y., Li Q, & Wang L. (2020). Chromodomain-helicase-DNA-binding protein 1-like (CHD1L) silencing inhibits gastric cancer cell proliferation, invasion, and migration. Translational Cancer Research, 9(11), 6660-6671.

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Live-Cell Imaging of Cytoskeleton Dynamics

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For live-cell imaging, 35-mm glass-bottom dishes (MatTek Corp.) were coated with 10 µg/ml fibronectin (PHE0023; Gibco) in PBS for ≥3 h at 37°C, washed with PBS twice, and immersed in complete DMEM without phenol red (01-053-1A; Biological Industries) before seeding of cells. For labeling actin and microtubule cytoskeleton, cells were incubated with 0.2 µM SiR-Actin (CY-SC001; Cytoskeleton) and SiR-Tubulin (CY-SC002; Cytoskeleton) for 6 h, respectively.
Time-lapse images of cells with transient transfection were acquired with the Olympus CellSens Dimension system, consisting of an Olympus SpinSR10 Ixplore spinning disk confocal microscope and a Yokogawa CSU-W1 confocal scanner. Appropriate filters, heated sample environment (37°C), controlled 5% CO₂, and UplanApo 100×/1.5 oil objective (Olympus Corp.) were used. The recording was set as every 10 min for 12 h, and one focal plane was recorded for all live-cell videos.

Fan C., Shi X., Zhao K., Wang L., Shi K., Liu Y.J., Li H., Ji B, & Jiu Y. (2022). Cell migration orchestrates migrasome formation by shaping retraction fibers. The Journal of Cell Biology, 221(4), e202109168.

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Elastic Micropillar Arrays for Cellular Traction

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Elastic micropillar arrays were fabricated in polydimethylsiloxane (PDMS). PDMS (Sylgard 184, Dow) was mixed in a 1:10 weight ratio according to the manufacturer specifications, poured on a silicon mould and cured at 60 °C for 1 h, resulting in PDMS with a spring constant k = 1.36 nN/µm. After curing, PDMS pillars were peeled-off the mould in phosphate buffered saline (PBS) and stored at 4 °C. Prior to seeding cells, PDMS pillars were coated with 10 μl/mL fibronectin (FN; Gibco, PHE0023) in PBS for 1 h at 37 °C. Cells were seeded on the FN-coated pillars and incubated for 1 h at 37 °C and 5% CO₂ before analysing them. Each sample was imaged at 37 °C for up to 30 min on an inverted microscope (Nikon Ti Eclipse, C-LHGFI HG Lamp, CFI Plan Fluor 40× NA 0.6 air objective) fitted with a Neo sCMOS camera (Andor, Oxford, UK) using NIS elements AR software. Each cell was recorded for 1 min with a frame rate of 1 frame/s. Data analysis was carried out with a custom MATLAB script to quantify pillar deflection and traction forces exerted on each pillar were calculated based on the deflection of the pillar and the spring constant.

Matellan C., Lachowski D., Cortes E., Chiam K.N., Krstic A., Thorpe S.D, & del Río Hernández A.E. (2023). Retinoic acid receptor β modulates mechanosensing and invasion in pancreatic cancer cells via myosin light chain 2. Oncogenesis, 12(1), 23.

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Cytoskeletal Reinforcement Dynamics

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Magnetic beads (4.5 µm, Dynabeads M-450, Thermo Fisher Scientific) were coated with fibronectin (Gibco, PHE0023) following the manufacturer’s instructions. Suit2 cells were incubated with fibronectin-coated beads for 30 min at 37 °C and then thoroughly washed with PBS to remove unbound beads. Individual cell-bound beads were then subjected to a pulsatile force regime using magnetic tweezers consisting of a 3 s, 6 nN force pulse, followed by a 4 s rest period, repeated for 12 pulses over ~100 s. The bead trajectories were recorded using an inverted microscope (Nikon Ti-Eclipse, C-LHGFI HG Lamp, CFI Plan Fluor 40× NA 0.6 air objective) fitted with a Neo sCMOS camera (Andor) with NIS elements AR software, and analysed using a custom MATLAB script. The amplitudes of each pulse were extracted from bead trajectories and normalised to the 1st pulse. The amplitudes of the 1st and 12th pulse were compared to quantify the decrease in amplitude of the bead movement as a result of cytoskeletal reinforcement.

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