Relevant antibodies were used to evaluate the expressions of S100A9, LDHA, PGK1, ENO1, β-catenin and c-Myc in tumour cells. The cells were centrifuged at 2800 RPM at 4 °C for 5 min, fixed with 4% paraformaldehyde and permeabilised with 0.1% Triton X-100 (BIOSS, P0096) at room temperature for 20 min. The cells were then washed with PBS (Solarbio, P1022) and blocked with 3% bovine serum albumin (BSA) (BIOSS, bs-0292P) at room temperature for 30 min. Then, incubation with primary antibodies (1:300 dilution) was performed at 4 °C overnight and HRP-tagged secondary antibody (1:500 dilution) for 50 min at room temperature. DAPI (BIOSS, C02-04002) was used for nucleus staining. Anti-fade mounting medium (Solarbio, S2110) was used for mounting.
P1022
Manufactured by Solarbio
Sourced in China
The P1022 is a laboratory equipment designed for research applications. It is a high-performance instrument that can be used for various experimental procedures. The core function of the P1022 is to provide precise measurements and data analysis capabilities to support scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
E672002, by Sangon (2 mentions)
Matrigel, by Corning (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Bioss Antibodies (1 mentions)
Bs 0292p, by Bioss Antibodies (1 mentions)
S2110, by Solarbio (1 mentions)
Annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis detection kit, by Beyotime (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
G1063, by Solarbio (1 mentions)
D7500, by Nikon (1 mentions)
Rnase a, by Merck Group (1 mentions)
Proteinase k, by Roche (1 mentions)
Jem 1200ex transmission electron microscope, by JEOL (1 mentions)
D8371, by Solarbio (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Ixplore pro, by Olympus (1 mentions)
E607309, by Sangon (1 mentions)
Transwell chamber, by Corning (1 mentions)
Crystal violet, by Sangon (1 mentions)
13 protocols using p1022
1
Immunofluorescence Analysis of Tumor Markers
2
Isolation and Characterization of UCMSC-Derived Exosomes
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When the isolated human UCMSCs at the fourth to sixth passage incubated in DMEM with 10% fetal bovine serum (FBS; FSP100, ExCell Biotech, Shanghai, China) reached 70%-80% confluence, the medium was centrifuged at 2,000 × g for 15 min at 4°C and then filtered with a 0.2 mm filter. Subsequently, the filtered medium was ultracentrifuged at 10,000 × g at 4°C. To remove the supernatant, it was ultracentrifuged at 100,000 × g for 20 min. After washing using the phosphate-buffered saline (PBS; P1022, Solarbio, Beijing, China), the exosomes derived from the human UCMSCs were stored at -80°C. The identification of exosomes was performed by measuring the surface marker using western blotting [33 (link)].
3
Quantifying ZIKV Entry in U87MG Cells
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U87MG cells were seeded in 24-well-plate at a density of 0.05−0.1 million per well and incubated with ZIKV at MOI = 1 for 4 h at 37 °C. The supernatant was then removed and the cells were washed three times with phosphate-buffered saline (PBS, Solarbio, P1022). Total RNA was extracted followed by qRT-qPCR to measure viral entry.
4
Annexin V-FITC/PI Apoptosis Detection
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The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection Kit (C1062M; Beyotime Institute of Biotechnology) was employed for the detection of cell apoptosis according to the instructions of manufacturers. In brief, SW480 and LOVO cells (1×104) were harvested and washed with phosphate-buffered saline (PBS; P1022; Beijing Solarbio Science & Technology Co., Ltd.) after relevant sevoflurane treatment and transfection. Then 5 µl Annexin V-FITC and 5 µl PI were added to stain cells for 15 min at room temperature in the dark. Finally, the rate of apoptotic cells was examined via a FACScan® flow cytometer (BD Biosciences) within 1 h and analyzed with software FlowJo (7.6.1; FlowJo LLC). The apoptotic rate was calculated as the sum of the early apoptosis rate and the late apoptosis rate.
5
Cell Sorting with BD FACS Aria
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The Influx or BD FACS Aria (FACAria Fusion SORP) cell sorter was set with a 100 μm tip/nozzle and multiple lasers (355, 488, 561, and 647 nm) in the collection mode. 0.4 X PBS was prepared as the sheath liquid by diluting 10 X PBS (Solarbio, P1022) with ultrapure water. The cells were sorted into 15 mL tubes at a flow rate of no more than 2. The number of sorted cells was set to meet the experimental requirements.
6
Colorimetric Assay for HepG2 Cell Viability
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HepG2 Cells (2000 cells per well) were seeded in 35 mm plates and incubated at 37 °C [43 (link)]. After overnight incubation, cells were treated with 200 μM I3C for 12 h. After replacing the medium, cells were incubated for 10 days. Plates were rinsed with phosphate-buffered saline (PBS, P1022, Solarbio, Beijing, China) and 2 mL of fixation reagent methanol was added for 15 min. Next, the cells were washed with PBS again and stained with 0.1% crystal violet (G1063, Solarbio, Beijing, China) for 10 min at room temperature. Plates were imaged using a digital camera (D7500, Nikon, Tokyo, Japan).
7
Extraction of Genomic DNA from Cells
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Cells were collected by centrifugation at 4°C for 3 min at 300 ×g, washed with 1× PBS (P1022, Solarbio), and resuspend with 600 μl Lysis Buffer (10 mM Tris–HCl, pH 8.0; 10 mM EDTA, pH 8.0; 10 mM NaCl; 0.5% SDS; 50 ng/μl RNase A (R6513, Sigma); 1 μg/μl Proteinase K (3115852001, Roche)). The suspension was incubated at 55°C for 40–60 min with interval mixing. gDNA was extracted by phenol:chloroform:isoamyl alcohol (P3803, Sigma) extraction and ethanol precipitation, and stored at −80°C.
8
Exosome Morphology Analysis by TEM
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Exosome pellets were suspended in PBS (P1022; Beijing Solarbio Science & Technology Co., Ltd.) and then fixed with 4% paraformaldehyde (E672002; Shanghai Sangon Biotech Co., Ltd.) overnight at 4°C and 4% glutaraldehyde (A600875; Shanghai Sangon Biotech Co., Ltd.) for 30 min at 4°C in phosphate buffer (pH 7.4), and maintained overnight at 4°C until the TEM assay. The exosomes were placed on a 400-mesh carbon-coated copper grid and stained with 2% phosphotungstic acid solution (pH 7.0; G1599; Solarbio) for 2 min at room temperature. The morphologies of the samples were examined with a JEM-1200EX transmission electron microscope (JEOL, Ltd.) at the magnification of ×50,000.
9
Adhesion Assay for Monocyte-Endothelial Interaction
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One day prior to the adhesion assay, 1 mL of HUVECs was seeded into 12-well plates at a density of 5 × 105 cells/mL and cultured in 5% CO2 at 37 °C until the cells reached 90% confluence. In the meantime, 1 mL of THP-1 cells (1 × 106 cells/mL) was stained with 1 μL of the fluorescent probe BCECF-AM (5 mM) (Beyotime, Shanghai, China) in the dark for 30 min. Then, the cells were washed twice using phosphate-buffered saline (PBS, pH = 7.4, P1022, Solarbio). After being resuspended, 1 mL (5 × 105 cells/mL) of the stained THP-1 cells was added to the monolayer formed by the HUVECs. After the cells were incubated at 37 °C for 60 min, the nonadherent cells were washed away using PBS. Under an inverted fluorescence microscope, five fields per well were randomly selected and photographed at 100 × magnification to calculate the number of adherent THP-1 cells. Image-Pro Plus (v 6.0) software was used for image analysis.
10
Orientin's Effects on RA-FLS Inflammation
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Orientin was brought from Med Chem Express (HY-N0405; MCE, USA) and was dissolved in dimethylsulfoxide (DMSO; D8371; Solarbio, China). Tumor necrosis factor alpha (TNF-α; HY-P7058; MCE) was dissolved in phosphate buffer saline (PBS; P1022; Solarbio). The solution was administered at 24 h after seeding of human RA-FLS. The cells were divided in nine groups: control (untreated group), orientin (5 μM) group, orientin (10 μM) group, orientin (30 μM) group, orientin (60 μM) group, TNF-α group, TNF-α + orientin (10 μM) group, TNF-α + orientin (30 μM) group, and TNF-α + (60 μM) group.
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