The patients' medical histories were ascertained by a questionnaire. Vigorous physical activity and smoking were avoided for at least 30 minutes before the measurement of the exercise capacity and the HD session. Blood was drawn from an arteriovenous shunt just before starting the HD sessions to determine hemoglobin, serum albumin, blood urea nitrogen, creatinine (Cr), uric acid, calcium, phosphate, lipids (high‐ and low‐density lipoprotein cholesterol, and triglycerides), and C‐reactive protein; values were analyzed at commercially available laboratories (Daiichi Pure Chemicals, Tokyo, Japan and Wako Pure Chemical Industries, Osaka, Japan). Serum carnitine fraction levels were determined as described previously.12 Serum interleukin‐6 (R&D Systems, Minneapolis, MN, USA), fibroblast growth factor‐21 (FGF‐21) (R&D Systems), myostatin (Immundiagnostik AG, Bensheim, Germany), and decorin (Abcam plc, Cambridge, UK) were determined by enzyme‐linked immunosorbent assay according to the manufacturer's instruction. Changes of all data both before and after treatment were calculated using the following formula: (post data − pre data)/pre data × 100 (%).
Decorin
Manufactured by Abcam
Sourced in United States
Decorin is a proteoglycan protein found in the extracellular matrix. It is involved in the regulation of collagen fibril assembly and organization.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 880, by Zeiss (2 mentions)
Fgf21, by R&D Systems (1 mentions)
Serum interleukin 6, by R&D Systems (1 mentions)
Cytokeratin 14, by Abcam (1 mentions)
Fibronectin, by Abcam (1 mentions)
Fluorophore conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Fabp4, by Abcam (1 mentions)
Pdgfrα, by R&D Systems (1 mentions)
α sma, by Abcam (1 mentions)
N cadherin, by Merck Group (1 mentions)
Integrin αv, by Abcam (1 mentions)
α1 catenin, by Abcam (1 mentions)
Dlk 1, by Abcam (1 mentions)
Lyve 1, by Abcam (1 mentions)
F4 80, by Abcam (1 mentions)
Fsp 1, by Abcam (1 mentions)
Epcam, by Abcam (1 mentions)
Radioimmunoprecipitation assay buffer, by Abcam (1 mentions)
Ecl western blotting detection reagent, by Thermo Fisher Scientific (1 mentions)
Mmp 2, by Merck Group (1 mentions)
9 protocols using decorin
1
Hemodialysis Protocol for Biomarker Analysis
2
Comprehensive Immunohistochemical Analysis of Tissue Samples
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The following antibodies were used: N-cadherin (clone GC4, Sigma-Aldrich C3865, 1:100 dilution), FSP1 (Abcam ab58597, 1:100 dilution), PDGFRα (R&D Systems AF1062, 1:50 dilution), Collagen I (Rockland 600–401–103–0.1, 1:150 dilution), Collagen III (Abcam ab7778, 1:150 dilution), Fibronectin (Abcam ab23750, 1:200 dilution), α-SMA (Abcam ab5694, 1:100 dilution), CD31 (Novus Biologicals NB100-2284, 1:500 dilution), EpCAM (Abcam ab92382, 1:500 dilution), Lyve-1 (Abcam ab14917, 1:500 dilution), Myf-5 (clone C-20, Santa Cruz sc-302, 1:500 dilution), CD45 (clone IBL-3/16, Abcam ab23910, 1:500 dilution), FABP4 (clone EPR3579, Abcam ab92501, 1:500 dilution), F4/80 (Abcam ab90247, 1:500 dilution), Cytokeratin 14 (clone EPR17350, Abcam ab181595, 1:200 dilution), Integrin αv (clone EPR16800, Abcam ab179475, 1:500 dilution), α1-catenin (clone EP1793Y, Abcam ab51032, 1:500 dilution), decorin (Abcam ab175404, 1:500 dilution), DLK-1 (Abcam ab21682, 1:200 dilution) and Ki67 (clone SP6, Abcam ab16667, 1:500 dilution). Fluorophore-conjugated secondary antibodies were purchased from Thermo Fisher Scientific (1:500 dilution). Exherin (ADH-1) was from TargetMol (T2637).
3
Protein Expression Analysis of hDF Hydrogels
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The hDF were loaded into hydrogels and collected based on similar methods to those described above. The collected cells were then washed with cold phosphate-buffered solution and treated with radioimmunoprecipitation assay buffer (Abcam, Cambridge, UK) and cold centrifuged a 14,000 rpm for 20 min. A Bradford protein assay from Bio-Rad, Richmond, CA, USA was used to evaluate for the levels of the different proteins. According to the manufacturer’s instructions, SDS-PAGE was used to separate the proteins, which were subsequently transferred onto polyvinylidene difluoride membranes. Target primary antibodies (β-actin, Abcam; MMP2, Millipore, Billerica, MA, USA; MMP9, Abcam; Decorin, Abcam) were placed onto the membranes and incubated overnight. Then, the membranes were washed and incubated with either horseradish peroxidase-conjugated anti-rabbit IgG (1:2,000 dilution; Genetex, Hsinchu, Taiwan) or horseradish peroxidase-conjugated anti-mouse IgG (1:2,000 dilution; Genetex) for 1 h at room temperature. The Fusion-Solo chemiluminescence system (Vilber, Paris, France) and ECL Western blotting Detection Reagents (Thermo Fisher Scientific, Waltham, MA, USA) were then used to detect the signals emitted from the samples.
4
Plasma Biomarker Quantification Protocol
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Blood was separated via centrifugation and plasma samples were aliquoted, flash frozen in liquid nitrogen, and stored at −80 °C until further analysis. Plasma concentrations of IL-6 were determined using Simple Plex cartridges run on the Ella platform (ProteinSimple, San Jose, CA) according to manufacturer’s instructions. High sensitivity ELISA kits were used to determine the plasma concentrations of myostatin (R&D Systems, Minneapolis, MN) and decorin (Abcam, Cambridge, United Kingdom) according to manufacturer’s instructions.
5
Histological Profiling of Tissue Samples
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Samples were fixed with 10% neutral buffered formalin (NBF) and embedded into paraffin blocks. Samples were stained with hematoxylin and eosin (H&E), Gomori’s trichrome (Thermo Fisher Scientific), and immunohistochemistry using antibodies against H2A.J (H2AFJ) (Origene, Rockville, MD), collagen IV (Rockland, Limerick, PA), decorin (Abcam, Cambridge, MA), elastin (Elastin Products Company, Owensville, MO), claudin 1 (Abcam), filaggrin (Abcam), transglutaminase (Proteintech, Rosemont, IL), and perilipin 3 (R&D Systems, Minneapolis, MN). Immunohistochemistry was performed using Leica automated stainer BOND-III (Leica, Buffalo Grove, IL) followed by image capture with digital image scanner NanoZoomer (Hamamatsu, Bridgewater, NJ).
6
Enzyme-Linked Immunosorbent Assay for ECM Binding
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Ninety-six well Enzyme-linked immunosorbent assay (ELISA) plates (Greiner Bio-One, Monroe, NC, USA) were coated with recombinant human ECM proteins (10 μg/mL): fibronectin (MilliporeSigma), decorin (Abcam, Cambridge, MA, USA), collagen I (MilliporeSigma), collagen II (MilliporeSigma), collagen III (MilliporeSigma), or collagen IV (MilliporeSigma); or 1 μg/mL of recombinant murine TNFα (PeproTech, Princeton, NJ, USA) in PBS overnight at 37 °C, followed by blocking with 2% BSA in PBS with 0.05% Tween 20 (PBS-T) for 1 h at room temperature. Then, wells were washed with PBS-T and further incubated with test antibody (100 μg/mL each) for 1 h at room temperature. After three washes with PBS-T, wells were incubated for 1 h at room temperature with horseradish peroxidase (HRP)-conjugated antibody against rat IgG (Jackson ImmunoResearch, Westgrove, PA, USA). After washes, bound antibodies were detected with tetramethylbenzidine substrate by measurement of absorbance at 450 nm with the subtraction of absorbance at 570 nm.
7
SMAD4, Versican, and Decorin Expression in TAAD
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Protein was extracted from human aorta tissues of 12 TAAD patients or SMCs culture medium, and subjected to western blot with mouse monoclonal antibodies against SMAD4, rabbit polyclonal antibodies against Versican or Decorin (1:1000, Abcam, Cambridge, UK). β-actin was used as internal control. The grey scales of target proteins were normalized against β-actin using Gel-Pro analyzer 4.0 (Media Cybernetics, Silver Spring, MD, USA).
8
Immunohistochemical Analysis of Periodontal Tissues
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Human periodontal soft tissue biopsies were incubated with primary antibodies diluted in 3% BSA/PBS overnight at 4℃. Primary antibodies used include Decorin (1:50, ab175404, Abcam), Osteoglycin (1:50, sc-374463, Santa Cruz), HLA-DR (1:50, ab92511, Abcam), and CD31 (1:50, ab9498, Abcam). Next day samples were incubated in Alexa-fluor 488, 594 Goat anti-Mouse or Goat anti-Rabbit secondary antibodies (Jackson Immunoresearch). Nuclei were counterstained with DAPI. Pictures were acquired using Zeiss LSM 880.
9
Immunohistochemical Analysis of ECM Proteoglycans
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Tissue specimens (tumour and non-tumour) were fixed in 10% formalin following which paraffin-fixed tissue-embedded blocks were prepared. Special 3-Aminopropyltriethoxysilane (APES)-coated slides were used for mounting the microtomised tissue specimen. Antigen retrieval was done using citrate buffer (pH 6.0). Washing was performed in Trisbuffered saline (TBS). Primary antibodies for Decorin (mouse monoclonal), Biglycan (mouse monoclonal) and Lumican (rabbit polyclonal) were purchased from Abcam (USA). All the primary antibodies were diluted in TBS at a concentration of 1:150 for both Decorin and Biglycan and 1:300 for Lumican. Anti-mouse and/or anti-rabbit secondary antibodies raised in goat were then added to the tissue section followed by treatment with avidin-conjugated horseradish peroxidase (HRP). After washing, each tissue section was then covered by freshly prepared diaminobenzidine (DAB) chromogen for few minutes until optimum development of brown-coloured peroxidase reactant product forms. Slides were counterstained by haematoxylin. Finally, slides were mounted after dehydrating with alcohol and acetone. The tissue expression was quantified by using immunoreactive score (IRS). 35
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