Human/Mouse Beclin 1 Antibody

Catalog #: AF5295 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF5295
AF5295-SP

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Detection of Human/Mouse Beclin 1 by Western Blot.

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Human/Mouse Beclin 1 Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human and mouse Beclin 1 in Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Beclin 1
Met12-Gln239
Accession # Q14457

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunocytochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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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.

Immunocytochemistry

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

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.

Long Name

Beclin1, Autophagy-related

Entrez Gene IDs

8678 (Human); 56208 (Mouse); 114558 (Rat)

Alternate Names

ATG6 autophagy related 6 homolog; ATG6; beclin 1 (coiled-coil, moesin-like BCL2 interacting protein); beclin 1 (coiled-coil, moesin-like BCL2-interacting protein); Beclin 1; beclin 1, autophagy related; beclin1; beclin-1; BECN1; Coiled-coil myosin-like BCL2-interacting protein; GT197; Protein GT197; VPS30