Cell culture reagents and supplies including Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12), trypsin and Antibiotic-Antimycotic (AbAm) were purchased from Invitrogen (Carlsbad, CA). Charcoal stripped fetal bovine serum (FBS) was from Sigma-Aldrich (St. Louis, MO). α7nAChR antibody was from Novus Biologicals (Littleton, CO). Other antibodies for α4nAChR, β2nAChR, caldesmon, α-SMA, calponin-1 and transgelin/SM-22 were obtained from Abcam (Cambridge, UK). β-actin antibody was from ABM Biological Materials (Richmond, Canada). Pro-inflammatory cytokines, tumor necrosis factor alpha (TNFα) and interleukin-13 (IL-13) were from R&D Systems (Minneapolis, MN). RIPA cell lysis/extraction buffer and A/G PLUS Agarose beads for co-immunoprecipitation techniques were obtained from Thermo Fisher Scientific (Waltham, MA) and SantaCruz Biotechnology (Dallas, TX) respectively. Pharmacological inhibitors for signaling pathways NFkB (SN-50) and STAT6 (AS1517499) were from SantaCruz Biotechnology (Dallas, TX) and Millipore Sigma (Burlington, MA) respectively. STAT3 (SD 1008), AP1 (SR 11302), CREB (666-15) and MAPK (PD 98059) were from Tocris Bioscience (Bristol, UK). Fluorescent secondary antibodies were from Li-Cor Biosciences (Lincoln, NE). Other chemicals/drugs/antibodies were from Sigma-Aldrich unless otherwise specified.
Caldesmon
Manufactured by Abcam
Caldesmon is a protein that regulates the interaction between actin and myosin in smooth muscle cells. It is involved in the contractile function of these cells.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Fluorescent secondary antibody, by LI COR (1 mentions)
Charcoal stripped fetal bovine serum, by Merck Group (1 mentions)
Calponin 1, by Abcam (1 mentions)
α sma, by Abcam (1 mentions)
Interleukin 13 il 13, by R&D Systems (1 mentions)
Tropomyosin, by Merck Group (1 mentions)
Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Vimentin, by Merck Group (1 mentions)
α tubulin, by Merck Group (1 mentions)
Trans blot semi dry transfer cell, by Bio-Rad (1 mentions)
Fluorchem fc2 system, by Cell Biosciences (1 mentions)
Nupage novex 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions)
α sma, by Merck Group (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
2 protocols using caldesmon
1
Cell culture protocol for nAChR signaling
2
Western Blot Analysis of Cell Markers
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Cells were lysed in a cold RIPA buffer (Thermo Scientific) containing Halt protease and phosphatase inhibitor cocktails (Thermo Scientific). Protein concentrations were determined using a Pierce BCA Protein Assay Kit (Thermo Scientific). Equal amounts of cell lysates were then separated on a NuPAGE Novex 4–12% Bis-Tris gel (Life Technologies) and transferred to PVDF membranes using a Transblot Semi-Dry Transfer Cell (Biorad). Membranes were blocked with a 5% non-fat dry milk solution and then examined with primary antibodies against αSMA (1:2500, Sigma), caldesmon (1:10000, Abcam), vimentin (1:500, Sigma), E-cadherin (1:1000, Cell Signaling), tropomyosin (1:1000, Sigma) α-tubulin (1:1000, Sigma), and β-Actin (1:1000, Cell Signaling). Blots were imaged on a FluorChem FC2 system (Cell Biosciences) through horseradish peroxidase (HRP)-conjugated secondary antibodies (1:2000, Cell Signaling) and SuperSignal West Pico Chemiluminescent Substrate (Pierce).
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