The contents of IL-1β and TNF-α in synovial fluid were determined by the ELISA method. The frozen synovial fluid was removed and centrifuged to obtain supernatant after being restored to room temperature. The samples were detected according to ELISA kit procedures (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and the absorbance values were measured at 450nm with an enzyme labelling instrument (Thermo Fisher Scientific, Waltham, USA). The contents of IL-1β and TNF-α were calculated by comparing them with the standard substance. Three wells were set for each index.
Enzyme labelling instrument
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Enzyme Labelling Instrument is a specialized piece of laboratory equipment designed to label enzymes with various tags or markers. It facilitates the process of conjugating enzymes with specific labels, enabling researchers to study and track the behavior of these enzymes in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using enzyme labelling instrument
1
Quantifying Inflammatory Cytokines in Synovial Fluid
2
Colon Tissue Protein Extraction and Analysis
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Proteins were extracted from the colon tissues using RIPA Lysis Buffer (Wuhan Servicebio Technology Co., Ltd, China) and the protein concentration was determined by bicinchoninic acid (BAC) method on the enzyme labelling instrument (Thermo Fisher Scientific, USA). The SDS-PAGE was used to separate the protein, and subsequently transferred to polyvinylidene fluoride (PVDF) membranes. The membranes were first blocked for 1h with 5% BSA at room temperature, and then probed with primary antibodies of MMP1 (Affinity, dilution of 1:1000), MMP3 (Abcam, dilution of 1:5000), MMP7 (Abcam, dilution of 1:1000), MMP9 (Abcam, dilution of 1:1000), MMP12 (Proteintech, dilution of 1:1000) and β-Actin (Affinity, dilution of 1:5000) at 4°C for overnight and then incubated with HPR-conjugated antibody at 37°C for one hour. Finally, the protein bands were recorded by ELC (Thermo Fish Scientific, USA) and quantified by Image J software to evaluate the protein relative expressions compare with β-Actin.
3
Characterization of Nanofibrous Mat Annealing
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The phase and surface information of nanofibrous mats before and after annealing treatment was characterized through XRD and FT-IR. The surface topography and structure of the samples were observed using field-emission scanning electron microscope (FESEM, S4800, Hitachi, Japan) and high-resolution transmission electron microscope (HRTEM, Hitachi, Japan). X-ray photoelectron spectroscopy (XPS) characterization was performed on an ESCALAB250Xi (Thermo Fisher Scientific, USA) spectrometer using Al Kα radiation as the excitation source. The absorbance of MB solution was detected by enzyme labelling instrument (Thermo Fisher Scientific, USA).
4
Quantifying Spinal Cord Inflammatory Markers
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Mice were deeply anaesthetized with intraperitoneal injection of 1% pentobarbital sodium (50 mg/kg) followed by immediate perfusion with 30 mL of precooled saline through the aorta. The spinal cords on the injured side, including the lumbar enlargement and approximately 1 cm in length, were obtained. Protein extraction was performed as follows. The obtained spinal cords were placed into a centrifuge tube, and RIPA lysis buffer and protease inhibitor were added. The samples were homogenized intermittently for 4 min on ice and centrifuged at 1000 rpm at 4 °C for 15 min. The precipitate was discarded, and the supernatant was transferred to another centrifuge tube. Then, the protein concentrations in the samples were analysed with a BCA kit. ELISA was performed according to the directions of the kits. The absorbance was measured at 450 nm with an enzyme labelling instrument (Thermo Fisher Scientific, Waltham, USA), and standard curves were constructed. According to the sample OD value, the levels of IL-1β, IL-6 and TNF-α in the spinal cord were calculated.
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