Smoothelin

Manufactured by Abcam

Sourced in United Kingdom

Smoothelin is a cytoskeletal protein found in smooth muscle cells. It plays a role in the regulation of smooth muscle contraction.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Mouse anti-smoothelin (2 citations)

4 protocols using smoothelin

Investigating VEGF-A Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

Recombinant human VEGF-A protein was obtained from R&D Systems and S-Nitrosoglutathione (GSNO) was obtained from santa cruz. Antibodies to pVEGFR2, VEGFR2, p STAT1, STAT1, p STAT3, STAT3, pSTAT5 and STAT5 were purchased from Cell Signaling; ACTA2, SM22, Myh11 and Smoothelin antibodies were purchased from Abcam; Myc antibody was purchased from proteintech.

Liao X.H., Xiang Y., Li H., Zheng D.L., Xu Y., Xi Yu C., Li J.P., Zhang X.Y., Xing W.B., Cao D.S., Bao L.Y, & Zhang T.C. (2017). VEGF-A Stimulates STAT3 Activity via Nitrosylation of Myocardin to Regulate the Expression of Vascular Smooth Muscle Cell Differentiation Markers. Scientific Reports, 7, 2660.

+ Open protocol

+ Expand

Smooth Muscle Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

PRF was obtained from Anhui Joyfar Pharmaceutical Co., Ltd, (Bozhou, China), while Dulbecco’s modified Eagle’s medium (DMEM)/F12 and fetal bovine serum (FBS) were purchased from Life Technologies (Carlsbad, CA, USA). Dimethyl sulfoxide (DMSO), MTT and recombinant human PDGF-BB were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse anti-cyclin D1, proliferating cell nuclear antigen (PCNA), cyclin-dependent kinase (CDK) 4, α-SMA, desmin, smoothelin, phospho-protein kinase B (Akt), total Akt, phospho-ERK, total ERK, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies and rabbit anti-mouse secondary antibodies were obtained from Abcam (Cambridge, UK).

LI H., LUO K, & HOU J. (2014). Inhibitory effect of Puerariae radix flavones on platelet-derived growth factor-BB-induced proliferation of vascular smooth muscle cells via PI3K and ERK pathways. Experimental and Therapeutic Medicine, 9(1), 257-261.

+ Open protocol

+ Expand

Immunofluorescence Staining of Endothelial Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

CD31, α-SMA (Sigma) and smoothelin (Abcam) primary antibodies (Sigma) were diluted (1∶400) in blocking buffer. 100 μl of the diluted antibody was added to each of the wells (1 h; 37°C). After washing with blocking buffer, 100 μl of AlexaFluor594-conjugated goat-anti-mouse secondary antibody was added for 1 h at 37°C (1∶400; Molecular Probes, Darmstadt, Germany). The cell nuclei were counterstained using DAPI nucleic acid stain (Molecular Probes). As negative controls, samples were incubated only in diluent and secondary antibody. Endothelial cells served as a positive control. All stained wells were viewed using an epifluorescence microscope (AxioObserver; Carl Zeiss). Images were acquired using a monochromatic digital camera (MRm; Carl Zeiss) and processed using digital software (AxioVision 4.8; Carl Zeiss).

Weinandy S., Babczyk P., Dreier A., Unger R.E., Flanagan T.C., Kirkpatrick C.J., Zenke M., Klee D, & Jockenhoevel S. (2014). Ovine Carotid Artery-Derived Cells as an Optimized Supportive Cell Layer in 2-D Capillary Network Assays. PLoS ONE, 9(3), e91664.

+ Open protocol

+ Expand

Protein Expression Analysis of Smooth Muscle Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tissues were lysed with RIPA buffer (100mM Tris (pH 7.4) (Sigma Aldrich), 4M Urea (Sigma Aldrich), 5mM EDTA (Sigma Aldrich), 0.5% SDS (Sigma Aldrich), 0.5% Nonidet P-40 (Sigma Aldrich), and protease inhibitor cocktail (Complete mini, Roche)) 36h after seeding. 15μg of total protein for each sample was run through a 4–15% Tris-HCL gel (Bio-Rad, Hercules, CA) at 120V for 2h. Then the samples were transferred to 0.2μm PVDF membrane (Bio-Rad). The following primary antibodies were incubated overnight at 4°C: smoothelin (Abcam), Calponin (Santa Cruz Biotechnology, Dallas, TX), Caldesmon (Santa Cruz Biotechnology), and β-actin (Santa Cruz Biotechnology). Protein bands were visualized after 1h incubation in LICOR secondary antibodies on the LICOR Odyssey (LICOR, Lincoln, NE). Image Studio Lite Version 3.1 was used to quantify protein amounts via densitometry. Each protein band value was normalized to its respective β-actin loading control and then normalized to the 10kPa substrate construct.

Steucke K.E., Tracy P.V., Hald E.S., Hall J.L, & Alford P.W. (2015). Vascular smooth muscle cell functional contractility depends on extracellular mechanical properties. Journal of biomechanics, 48(12), 3044-3051.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!