Detection of Human and Rat Coactosin-like Protein 1/COTL1 by Western Blot.
Western blot shows lysates of JAR human choriocarcinoma cell line, NCI‑H345 human small cell lung carcinoma cell line, human placenta tissue, and rat placenta tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat Coactosin-like Protein 1/COTL1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7865) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Coactosin-like Protein 1/COTL1 at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Coactosin-like Protein 1/COTL1 in NCI‑H128 Human Cell Line.
Coactosin-like Protein 1/COTL1 was detected in immersion fixed NCI‑H128 human small cell lung carcinoma cell line using Sheep Anti-Human/Mouse/Rat Coactosin-like Protein 1/COTL1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7865) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 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: Coactosin-like Protein 1/CotL1
COTL1 (Coactosin-like Protein) is both a cytoplasmic and plasma-appearing 15-16 kDa member of the coactosin subfamily, ADF/Actin Depolymerizing Factor family of actin-binding proteins. It is widely expressed, and found in cell types such as neutrophils, and tissues such as placenta, lung and kidney. Functionally, COTL1 interacts noncovalently with both F-actin and 5-lipoxygenase/5LO. These interactions appear to be mutually exclusive. A COTL1:F-actin interaction leads to actin binding without actin polymerization, while a 5LO:COTL1 interaction has two potential outcomes; first, 5LO sequesters COTL1, leading to a failure of actin binding, and second, COTL1 can serve as a scaffold for 5LO activity, facilitating the production of either 5HPETE or LTA4. Human COTL1 is 142 amino acids (aa) in length. It is principally composed of one ADF-H domain (aa 2-130) that possesses a utilized phosphorylation site at Ser115, and two acetylation sites at Lys102 and Lys126. COTL1 may form noncovalent homodimers and oligomers, but not when complexed to F-actin. There is one potential isoform variant that shows a 106 aa substitution for aa 1-53. Full-length human and mouse COTL1 share 95% aa sequence identity.
Entrez Gene IDs:
23406 (Human); 72042 (Mouse); 361422 (Rat)
CLP; CLPFLJ43657; Coactosin like Protein 1; coactosin-like 1 (Dictyostelium); Coactosin-like Protein 1; coactosin-like protein; COTL1; MGC19733
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