Detection of Human Pref‑1/DLK1/FA1 by Western Blot.
Western blot shows lysates of human adrenal gland tissue and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Pref‑1/DLK1/FA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1144) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Pref‑1/DLK1/FA1 at approximately 45-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Pref‑1/DLK1/FA1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human Pref‑1/DLK1/FA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1144) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
Pref-1 (Preadipocyte factor 1; also DLK-1 and FA1) is a 58-65 kDa member of the Notch/Serrata/Delta family of proteins. It is expressed in prechondrocytes and preadipocytes and appears to block progenitor cell differentiation into mature cell lineages. Mature human Pref-1 is a 360 amino acid (aa) type I transmembrane N- and O-linked glycoprotein. It contains a 280 aa extracellular region (aa 24-303), a 24 aa transmembrane segment (aa 304-327), and a 56 aa cytoplasmic domain (aa 328-383). The extracellular region contains six EGF-like domains, and undergoes proteolytic cleavage to generate a bioactive 50 kDa fragment, plus three 25‑31 kDa fragments that show no activity. There are multiple potential splice variants. One shows a deletion of aa 229-301, a second possesses a six aa substitution for aa 1‑52, a third shows a deletion of aa 210-277, while a fourth contains a six aa substitution for aa # 207-383. Over aa 24-297, human Pref-1 shares 82% aa identity with mouse Pref-1.
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