Total lysates of human OA chondrocytes were prepared using RIPA buffer (Biosesang, Gyeonggi-do, Korea). Primary antibodies against MMP3, MMP13 (1:1000 dilution, Cell Signaling Technology, Danvers, MA, USA), COL2A1 (1:1000 dilution, Abcam), ACAN (1:500 dilution, Invitrogen), p38, p-p38, ERK, p-ERK, JNK, p-JNK, p65, p-p65, IκBα, p-IκBα, or β-actin (1:1000 dilution, Cell Signaling Technology) and appropriate HRP-conjugated secondary antibodies (1:2000 dilution, Bethyl) were used. Enhanced chemiluminescence (ECL) detection kit (Thermo Fisher Scientific) was used to detect for specific bands. Chemiluminescent signals were analyzed on a ChemiDoc XRS gel imaging system (Bio-Rad Laboratories, Hercules, CA, USA), and intensity of specific bands was quantified using Image J software (NIH, MD, USA).
Enhanced chemiluminescence ecl detection kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Enhanced chemiluminescence (ECL) detection kit is a laboratory equipment used for detecting and quantifying protein expression levels in Western blot analysis. The kit contains reagents that generate a luminescent signal when combined with the target protein, allowing for the visualization and measurement of the protein of interest.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions)
Mmp13, by Cell Signaling Technology (1 mentions)
Col2a1, by Abcam (1 mentions)
P iκbα, by Cell Signaling Technology (1 mentions)
Hrp conjugated secondary antibody, by Fortis Life Sciences (1 mentions)
Chemidoc xrs gel imaging system, by Bio-Rad (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
Ripa buffer, by Biosesang (1 mentions)
Arbutin, by Merck Group (1 mentions)
α msh, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by Welgene (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay kit, by Promega (1 mentions)
Trypsin edta, by GenDEPOT (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Milli q purification system, by Merck Group (1 mentions)
Radio immunoprecipitation assay ripa buffer, by Thermo Fisher Scientific (1 mentions)
4 protocols using enhanced chemiluminescence ecl detection kit
1
Osteoarthritis Chondrocyte Protein Analysis
2
Melanoma Cell Proliferation Assay
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The B16F10 mouse melanoma cells (ATCC CRL-6475) were collected from the American Type Culture Collection (Manassas, VA, USA). Dulbecco’s modified Eagle’s medium (DMEM) was bought from WELGENE Inc. (Gyeongsan-si, South Korea). Arbutin, α-MSH, and dimethyl sulfoxide (DMSO) were from Sigma-Aldrich Co. (St. Louis, MO, USA). Fetal bovine serum (FBS), penicillin-streptomycin, and trypsin-EDTA were purchased from GenDEPOT Inc. (Barker, TX, USA). The CellTiter 96® AQueous One Solution Cell Proliferation Assay Kit was collected from Promega (Madison, WI, USA). The BCA Protein Assay Kit, Enhanced Chemiluminescence (ECL) Detection Kit, and NE-PER nuclear and cytoplasmic extraction reagents were from Thermo Scientific (Rockford, IL, USA). Thee cAMP Assay Kit was purchased from R&D Systems, Inc. (Minneapolis, MN, USA).
3
Purified MAN Modulation of LPS-Induced Effects
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MAN with the purity of 98% (based on HPLC-UVD analysis) was brought from SanHerb Bioscience Inc. (Chengdu, Sichuan, China). Lipopolysaccharide (LPS, from the Gram-negative bacteria E. coli 055:B5), phosphate buffered saline (PBS), BCA protein quantitative kit, NAD, nicotinamide mononucleotide (NMN), HRP conjugated streptavidin, and HRP/biotin conjugated secondary antibodies were purchased from KeyGen Biotech (Nanjing, Jiangsu, China). Fetal bovine serum (FBS) and Dulbecco's Modified Eagle Medium (DMEM) were from TianHang Biotechnology (Hangzhou, Zhejiang, China). Enhanced chemiluminescence (ECL) detection kit, defatted milk powder, and bovine serum albumin (BSA) were from Thermo Scientific (Rockford, IL, USA). Primary antibodies used in both western blot and immunohistochemical assays were obtained from Affinity Biosciences (Cincinnati, OH, USA). Ultrapure water was prepared by using a Milli-Q purification system (Millipore, Bedford, MA, USA).
4
Investigating KLF3-AS1 and miR-338-3p in Osteosarcoma
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ScienCell provided the human normal osteoblast hFOB 1.19. OS cell lines HOS, Saos‐2, and SW1353 were obtained from American Type Culture Collection (ATCC). Dulbecco's modified Eagle medium (DMEM), fetal bovine serum (FBS), and the Lipofectamine 3000 were sourced from Invitrogen. KLF3‐AS1‐siRNA (si‐lnc), MEF2C‐siRNA (si‐MEF2C) and its negative control (si‐NC), small hairpin targeting KLF3‐AS1 (sh‐lnc) and its sh‐NC, miR‐338‐3p mimic, mimic‐NC, miR‐338‐3p inhibitor (inhibitor), and inhibitor‐NC were all acquired from Sangon Biotech. The CCK‐8 kit and dimethyl sulfoxide (DMSO) used for cell viability assays were procured from Aladdin. Crystal violet and RIP assay‐related kit were from Sigma Aldrich. Primary antibodies (MEF2C, cat.no. ab211493; GAPDH, cat.no. ab8245; Bcl‐2, cat.no. ab194583; and Bax, cat.no. ab32503), and HRP‐conjugated secondary antibody (cat.no. ab6789) were bought from Abcam. The radioimmunoprecipitation assay (RIPA) buffer, enhanced chemiluminescence (ECL) detection kit, and PARIS™ Kit were from Thermo Fisher Scientific. The Total RNA Extraction Kit was acquired from Promega; the First‐Strand cDNA Synthesis Kit was from APExBIO Technology; and from Qiagen, we acquired a SYBR Green FAST Mastermix Kit (Dusseldorf, GER). Four‐ to five‐week‐old female BALB/c nude mice weighing 20–28 g, from Esebio, were used to establish tumor xenograft models.
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