Calponin regulates smooth muscle cell contraction and is a marker of smooth muscle cell differentiation. Calponin, an Actin- and Tropomyosin-binding protein, is characterized as an inhibitory factor of smooth-muscle actomyosin activity. Calponin is implicated in the regulation of smooth muscle contraction through its interaction with F-Actin and inhibition of the Actin-activated MgATPase activity of phosphorylated Myosin. Both properties are lost following phosphorylation (primarily at Serine 175) by protein kinase C or calmodulin-dependent protein kinase II. The three forms of Calponin, Calponin 1 (basic Calponin), Calponin 2 (neutral Calponin) and Calponin 3 (acidic Calponin), are found in smooth muscle tissue. Additionally, Calponin 2 is found in heart muscle tissue and Calponin 3 is found in the brain.
Background References
1. Battiston KG et al. Monocyte/macrophage cytokine activity regulates vascular smooth muscle cell function within a degradable polyurethane scaffold. Acta Biomater 10:1146-55 (2014).
2. Nyp MF et al. TRIP-1 via AKT modulation drives lung fibroblast/myofibroblast trans-differentiation. Respir Res 15:19 (2014).
Sequence Similarity
Belongs to the calponin family.
Tissue Specificity
Smooth muscle, and tissues containing significant amounts of smooth muscle.
Western blot analysis of Calponin on different lysates with Rabbit anti-Calponin antibody (ET1606-17) at 1/1,000 dilution.
Lane 1: Mouse stomach tissue lysate Lane 2: Rat stomach tissue lysate
Lysates/proteins at 40 µg/Lane.
Predicted band size: 33 kDa Observed band size: 35 kDa
Exposure time: 3 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-17) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ICC staining of Calponin in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1606-17, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Calponin antibody (ET1606-17) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-17) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-Calponin antibody (ET1606-17) at 1/100,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-17) at 1/100,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Calponin antibody (ET1606-17) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-17) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-Calponin antibody (ET1606-17) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-17) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Western blot analysis of Calponin on different lysates with Rabbit anti-Calponin antibody (ET1606-17) at 1/1,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lysates/proteins at 20 µg/Lane. Exposure time: 53 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET1606-17, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 33 kDa Observed band size: 33 kDa
Western blot analysis of Calponin on different lysates with Rabbit anti-Calponin antibody (ET1606-17) at 1/1,000 dilution.
Lane 1: NIH/3T3 (Mouse fibroblast) cell lysate
Lysates/proteins at 20 µg/Lane. Exposure time: 53 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET1606-17, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 33 kDa Observed band size: 33 kDa
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