SalB and Rg1 were purchased from Shanghai Yousi Bio-Tech Co., Ltd (Shanghai, China). The structure of SalB (S1 Fig ) or Rg1 (S2 Fig ) was elucidated by 1H and 13C NMR spectrum using Bruker AM-400 spectrometer. Purity of SalB was 99.84%, and Rg1 was 99.78% evaluated by high performance liquid chromatography (S3 Fig ). Procarta cytokine profiling kit (Panomics, CA, USA), chloral hydrate (SCRC, Shanghai, China), 3,5-triphenyltetrazolium chloride (Sigma-Aldrich, USA), troponin-I antibody (abcam, Cambridge, United Kingdom), vimentin antibody (Calbiochem, Darmstadt, Germany), β-actin antibody (Cell-signaling, Boston, USA) and collagenase Type II (DingGuo, Shanghai, China) were commercially obtained. Mitochondrial membrane potential assay kit was purchased from Beyotime (Jiangsu, China). H9c2 cells were purchased from Cell bank of Chinese Academy of Sciences (Shanghai, China). All the culture reagents were products from Thermo Scientific (Beijing, China) unless specified otherwise.
Procarta cytokine profiling kit
Manufactured by Panomics
Sourced in United States
The Procarta cytokine profiling kit is a lab equipment product that enables the simultaneous detection and quantification of multiple cytokines in a single sample. The kit utilizes a multiplex bead-based assay technology to provide a comprehensive analysis of cytokine levels.
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Lab products found in correlation
Mitochondrial membrane potential assay kit, by Beyotime (1 mentions)
Am 400 spectrometer, by Bruker (1 mentions)
β actin antibody, by Cell Signaling Technology (1 mentions)
Troponin 1 antibody, by Abcam (1 mentions)
Bio plex cytokine assay system, by Bio-Rad (1 mentions)
Ebv ish kit, by Leica (1 mentions)
Luminex 200, by Bio-Rad (1 mentions)
6 protocols using procarta cytokine profiling kit
1
Evaluation of SalB and Rg1 in H9c2 Cells
2
Serum Survivin Concentration Measurement
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Serum survivin concentration was measured in archived frozen samples of the aforementioned prospective cohort study. Archived serum sample aliquots had been stored at –80°C and were thawed before use in the cytokine assay. The concentration of survivin, an antiapoptosis protein, was measured in serum using the Procarta cytokine profiling kit (Panomics, San Diego, CA, USA), and all measurements were performed in duplicate according to the manufacturer's instructions.
3
Multiplex Cytokine Profiling in Serum
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Serum samples were collected at diagnosis and stored at -80°C until analysis. Serum aliquots had not been previously thawed before use in the multiplex chemokine assay. We measured the levels of IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, and interferon-gamma (IFN-γ) in triplicate with the Procarta cytokine profiling kit (Panomics, CA, USA) using the Bio-Plex Cytokine Assay System (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer's instructions.
4
Quantifying Serum CCL3 and EBV Detection
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The serum CCL3 concentration was measured in archived frozen samples of the Samsung Medical Center prospective cohort study (NCT#00822731). Archived serum sample aliquots stored at −80°C were thawed for use in the cytokine assay. The CCL3 concentration was measured in serum using the Procarta cytokine profiling kit (Panomics, San Diego, CA, USA), and all measurements performed in duplicate.
EBER was detected using an EBV ISH kit (Leica Microsystems, Bannockburn, IL, USA). We employed EBV-negative lymphoid tissues and the hybridization mixture without EBV oligonucleotides as the negative control. A positive reaction was defined as >20% cells showing nuclear positivity.
EBER was detected using an EBV ISH kit (Leica Microsystems, Bannockburn, IL, USA). We employed EBV-negative lymphoid tissues and the hybridization mixture without EBV oligonucleotides as the negative control. A positive reaction was defined as >20% cells showing nuclear positivity.
5
Multiplexed Cytokine Profiling in Patients
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Serum aliquots were not thawed before we performed the multiplex cytokine assay. We measured the levels of regulated on activation, normal T cell-expressed and secreted (RANTES), IL-10, IL-6, interferon gamma-induced protein 10 (IP-10), and TNF-α with the Procarta cytokine profiling kit (Panomics, Fremont, CA) according to the manufacturer’s instructions, in duplicate. We also measured the level of plasminogen activator inhibitor 1 (PAI), a procoagulant parameter, with the same kit. Based on the cutoff value of each cytokine examined, patients were categorized into high and low groups. The optimal cutoff value for TNF-α was determined by receiver-operating characteristic (ROC) curve analysis. The area under the ROC curve was 0.66 (cutoff value, 0.285; 95% confidence interval [CI], 0.517 to 0.694). Because of the low sensitivity and specificity for other cytokines and PAI through ROC curve analysis, we used the median value as the cutoff point for PAI, RANTES, IL-10, and IP-10 analyses. Only 26% of all patients showed a positive value of cytokine IL-6, so we categorized the IL-6 level as detectable or undetectable.
6
Multiplex Cytokine Profiling in Cell Culture
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A Procarta cytokine profiling kit (Panomics, Santa Clara, CA, USA) was used according to the manufacturer's instructions to simultaneously detect 12 different human chemokines and cytokines in cell culture supernatants. Briefly, 50 μl of antibody beads was added to each well of the filter plate and washed with wash buffer. A volume of 50 μl of cell culture supernatant was then added to each well, incubated for at least 1 h at room temperature, and washed with wash buffer. Afterward, 25 μl per well of the Detection Antibody solution was added, and the filter plate was shaken at 500 r.p.m. for 30 min at room temperature. After adding streptavidin-PE, the signals were detected with a Luminex 200 instrument (Bio-Rad, Hercules, CA, USA). The chemokines and cytokines included in the present study were inflammation-associated factors and angiogenic cytokines (ICAM, IL-1β, IL-2, IL-6, IL-8, IL-10, IL-17A, IL-22, MCP-1, platelet-derived growth factor, stromal cell-derived factor (SDF)-1, and VEGFA). The results were based on three independently repeated experiments (Supplementary Table 1 ).
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