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Calponin

Manufactured by Agilent Technologies
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Sourced in Denmark

Calponin is a lab equipment product manufactured by Agilent Technologies. It is a protein that regulates smooth muscle contraction. The core function of Calponin is to inhibit the actin-myosin interaction, thereby modulating the contractile activity of smooth muscle cells.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Tween 20, by Merck Group (1 mentions) Calponin, by Abcam (1 mentions) α sma, by Agilent Technologies (1 mentions) Chamber slide, by Merck Group (1 mentions) Alexa fluor 488 labeled, by Abcam (1 mentions) Ventana benchmark xt, by Roche (1 mentions) Myogenin, by Roche (1 mentions) Sox10, by Biocare Medical (1 mentions) Factor xiiia, by Cell Marque (1 mentions) Ultra view polymer detection kit, by Roche (1 mentions) Desmin, by Agilent Technologies (1 mentions) β catenin, by BD (1 mentions) α sma, by Abcam (1 mentions) Cd31 antibody, by BD (1 mentions) Cellquest software, by BD (1 mentions) Facscan flow cytometer, by BD (1 mentions) α tubulin, by Merck Group (1 mentions) Integrin β1, by Santa Cruz Biotechnology (1 mentions) Fibronectin, by Merck Group (1 mentions)

Close spelling variants of this product

Anti-Calponin (4 citations) Calponin (clone CALP; (3 citations)

4 protocols using calponin

1

Immunocytochemical Characterization of Cell Populations

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To characterize and assess the purity of the obtained cell populations, immunocytochemistry was performed. Cells were cultured for 24–48 h on chamber slides (Millipore, Burlington, MA, United States) at a concentration of 20,000 cells/cm2 under the described culture conditions until a 90% confluence was obtained. Cells were fixed for 4 min in ice-cold acetone/methanol (1:1 v/v) and air dried for 10 min at RT. Slides were washed with phosphate buffered saline (PBS) with 0.1% Tween 20 (Sigma) and blocked with 10% goat serum in PBS prior to incubation with rabbit primary antibody against CD31 (Abcam, Cambridge, United Kingdom), Calponin (Abcam), vWF (Dako, Santa Clara, CA, United States), and α-SMA (Alexa Fluor 488 labeled, Abcam) overnight at 4°C. Anti-rabbit IgG (DAKO, Agilent, Santa Clara, CA, United States) was used as isotype control. Incubation of secondary Alexa Fluor 488 labeled goat-anti-rabbit (Dako) was performed for 1 h at RT for the CD31, Calponin, α-SMA, and isotype conditions. Counterstaining was performed with 1 μg/mL DAPI (Sigma) for 10 min at RT.
Oosterhoff L.A., Kruitwagen H.S., van Wolferen M.E., van Balkom B.W., Mokry M., Lansu N., van den Dungen N.A., Penning L.C., Spanjersberg T.C., de Graaf J.W., Veenendaal T., Zomerdijk F., Fledderus J.O., Spee B, & van Steenbeek F.G. (2019). Characterization of Endothelial and Smooth Muscle Cells From Different Canine Vessels. Frontiers in Physiology, 10, 101.
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2

Immunohistochemical Profiling of Rare Tumors

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Immunohistochemistry was performed on all cases, either at the time of original diagnosis or for the purposes this study. Whole-slide sections of formalin-fixed, paraffin embedded tumor tissue were cut at five-micron thickness, deparaffinized, and subjected to antigen retrieval using 10 mM citrate buffer at 92°C for 30 minutes. Immunohistochemistry was performed on all cases using antibodies for S100 (Ventana Medical Systems, Tucson, AZ), either Smooth Muscle Actin (Ventana) or Calponin (Dako, Carpinteria, CA), SOX10 (BioCare Medical, Concord, CA), Desmin (Dako), and β-catenin (BD Biosciences, San Jose, CA). In addition, for a subset of cases (n=6), PAX3 (Bioss Inc., Wolburn, MA) immunohistochemistry was also performed. Depending on tissue availability, immunohistochemistry was also performed in most cases using antibodies for myogenin (Ventana) and Factor XIIIa (Cell Marque, Rocklin, CA). All immunohistochemical signals were visualized using the Ultra view polymer detection kit (Ventana Medical Systems, Inc. Tucson, AZ) on a Ventana BenchmarkXT autostainer (Ventana). Staining was performed according to manufacturer's instructions in the presence of appropriate controls. “Focal” immunoreactivity was regarded as immunostaining in <10% of tumor cells.
Rooper L.M., Huang S.C., Antonescu C.R., Westra W.H, & Bishop J.A. (2016). Biphenotypic Sinonasal Sarcoma: an Expanded Immunoprofile Including Consistent Nuclear Beta-Catenin Positivity and Absence of SOX10 Expression. Human pathology, 55, 44-50.
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3

Characterization of Endothelial Progenitor Cells

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Fluorescence-activated cell sorting (FACS) analysis was performed by staining with 10 μl/ each of phycoerythrin (PE) or Fluorescein isothiocyanate (FITC) conjugated CD34 antibody (BD Biosciences), KDR/Flk-1 (VEGF receptor-2) antibody (R&D System Inc), VE-cadherin antibody (Bender MedSystem GmbH, Vienna, Austria), CD31 antibody (BD Biosciences), α-smooth muscle actin (α-SMA) antibody (R&D System Inc), Platelet-derived growth factor receptors (PDGFR-α) antibody, PDGFR-β antibody and CD45 antibody (BD Biosciences) for 30 min at 4°C in a dark room. Data were analyzed using a FACScan® flow cytometer and CellQuest® software (Becton Dickinson, Franklin Lakes, NJ). The ECFCs and SPCs were seeded onto an 8-well chamber glass microscope slide, and then were fixed in 1% paraformaldehyde and permeabilized with 0.1% Triton X-100. The cells were incubated with CD34 (SantaCruz, Dallas, TX), Von Willebrand factor (vWF), CD31, α-SMA (Abcam, Cambridge, UK), CD45 (BD Biosciences) and Calponin (DAKO, Glostrup, Denmark) antibodies for overnight at 4°C, and incubated with fluorescence-conjugated secondary antibodies for 1 hr at room temperature.
Phi J.H., Suzuki N., Moon Y.J., Park A.K., Wang K.C., Lee J.Y., Choi S.A., Chong S., Shirane R, & Kim S.K. (2017). Chemokine Ligand 5 (CCL5) Derived from Endothelial Colony-Forming Cells (ECFCs) Mediates Recruitment of Smooth Muscle Progenitor Cells (SPCs) toward Critical Vascular Locations in Moyamoya Disease. PLoS ONE, 12(1), e0169714.
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4

Immunofluorescence analysis of cell-cell junctions

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The following commercially available antibodies were used: claudin-5 (Zymed/Life Technologies), occludin (Transduction Laboratories, Franklin Lakes, NJ, USA), VE-cadherin (Cell Signaling, Danvers, MA, USA), N-cadherin (Transduction Laboratories), β1-integrin (Santa Cruz Biotechnologies, CA, USA), fibronectin (Sigma), calponin (DAKO, Glostrup, Denmark), SMA (Sigma), α-tubulin (Sigma), SRF (Santa Cruz Biotechnologies), phospho-Smad2/Smad3 (Cell Signaling), Smad2/3 (Cell Signaling). Peroxidase-conjugated anti-mouse and anti-rabbit secondary antibodies were obtained from Cell Signaling. Cy3-labeled anti-mouse and Cy3-labeled anti-rabbit antibodies were obtained from Jackson ImmunoResearch (Newmarket, UK). SB-431542 was purchased from Sigma. Y-27632 was obtained from Tocris (Bristol, UK). For inhibitor studies control cells were treated with vehicle.
Krizbai I.A., Gasparics Á., Nagyőszi P., Fazakas C., Molnár J., Wilhelm I., Bencs R., Rosivall L, & Sebe A. (2015). Endothelial-Mesenchymal Transition of Brain Endothelial Cells: Possible Role during Metastatic Extravasation. PLoS ONE, 10(3), e0119655.
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