Detection of Human/Mouse Beclin 1 by Western Blot.
Western blot shows lysates of T47D human breast cancer cell line, HeLa human cervical epithelial carcinoma cell line, CH1 mouse B cell lymphoma cell line, and A20 mouse B cell lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse Beclin 1 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF5295) followed by HRP‑conjugated Anti‑Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Beclin 1 at approximately 60 kDa (as indicated). This experiment was conducted using Immunoblot Buffer Group 2.
Beclin 1 in HeLa Human Cell Line.
Beclin 1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse Beclin 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5295) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (yellow; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to autophagosomes. 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.
Background: Beclin 1
Beclin 1 (Bcl-2 interacting coiled-coil protein; also know as ATG6) is a 60 kDa member of the Beclin family of proteins. It is part of a lipid-kinase complex that includes UVRAG and PI3 and induces autophagy; in the presence of Bcl-2, autophagy is repressed. Beclin 1 is widely expressed but abundantly found in mammary epithelium, glia and neurons. Human Beclin 1 is 450 amino acids (aa) in length and contains a Bcl-2 binding domain (aa 88‑150), a nuclear export signal motif (aa 180‑189), a coiled-coil region (aa 142‑270), and an evolutionary conserved domain (aa 244‑337). There is one potential alternate start site at Met88. Over aa 12‑239, human Beclin 1 is 98% aa identical to mouse Beclin 1.
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