Detection of Human, Mouse, and Rat Calponin 1 by Western Blot.
Western blot shows lysates of MDA‑MB‑231 human breast cancer cell line, MCF‑7 human breast cancer cell line, NMuMG mouse mammary gland epithelial cell line, and A7r5 rat thoracic aortic smooth muscle cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Calponin 1 Monoclonal Antibody (Catalog # MAB7900) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Calponin 1 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Calponin 1 in MDA‑MD‑231 Human Cell Line.
Calponin 1 was detected in immersion fixed MDA‑MD‑231 human breast cancer cell line using Mouse Anti-Human Calponin 1 Monoclonal Antibody (Catalog # MAB7900) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Calponin 1 in Human Small Intestine.
Calponin 1 was detected in immersion fixed paraffin-embedded sections of human small intestine using Mouse Anti-Human Calponin 1 Monoclonal Antibody (Catalog # MAB7900) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm of smooth muscle cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Calponin 1
CNN-1 (Calponin 1 [calcium and calmodulin-binding troponin T-like protein]; also Calponin basic, CaP and Calponin H1) is a 32-36 kDa cytoplasmic member of the calponin family of proteins. Although reportedly expressed in fibroblasts and endothelial cells, it actually appears to be restricted to smooth muscle and smooth muscle-like cells such as myoepithelium and myofibroblasts in the adult. CNN-1 interacts with F-actin in a phosphorylation-dependent manner. When nonphosphorylated, CNN-1 blocks actomyosin ATPase activity, contributing to the stabilization of actin stress fiber bundles. Thus, CNN-1 expression inhibits cell motility and the formation of podosomes. Human CNN-1 is 297 amino acids (aa) in length. It contains one CH/calponin homology domain (aa 30-127), and three consecutive calponin-like repeats (aa 164-268). The repeats are suggested to mediate actin binding. There are five potential Ser/Thr phosphorylation sites. Full-length human CNN-1 shares 97% aa sequence identity with mouse CNN-1.
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