|Twist‑1 in A549 Human Cell Line. Twist‑1 was detected in immersion fixed A549 human lung carcinoma cell line treated with 10ng/mL of Recombinant Human TGF beta 1 (Catalog # 240-B) for 48 hours using Sheep Anti-Human Twist‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6230) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (yellow, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
Intracellular Staining by Flow Cytometry
|Detection of Twist‑1 in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Sheep Anti-Human Twist‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6230, filled histogram) or control antibody (Catalog # 5‑001‑A, open histogram), followed by NorthernLights™ 493-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL012). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
Twist-1 (Twist-related protein 1; also H-Twist and bHLHa38) is a 25‑27 kDa class B member of the bHLH transcription factor family of proteins. It is widely expressed in embryo, and select adult cells such as white adipocytes. In fat, Twist-1 induces fatty acid oxidation via PGC-1 alpha and CPT-1, and promotes MCP-1 and TNF-alpha secretion by adipocytes. In epithelium, Twist-1 dysregulation represses E-Cadherin and induces N-Cadherin expression, resulting in an epithelial-to-mesenchymal transition that can lead to cancer. Human Twist-1 is 202 amino acids (aa) in length. It contains a bHLH domain (aa 109-164) with an embedded DNA-binding motif. Twist-1 forms homodimers, and heterodimerizes with TCF3, HAND 1 and HAND 2. Two distinct mutations exist that may impact its dimerization pattern. There is one seven aa insertion after Ile135, and a second seven aa insertion after Pro139. A third unrelated variant shows a four Gly insert after Gly92. Full-length human Twist-1 shares 98% aa identity with mouse Twist-1.
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