BECN1 Antibody

• Catalog Number: orb1239257
• Category: Antibodies
• Description: BECN1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, WB
• Predicted Reactivity: Bovine, Porcine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-Beclin-1 antibody (orb1239257) was raised against a peptide corresponding to 17 amino acids near the center of human Beclin-1. The immunogen is located between 40-90 amino acids of Beclin-1.
• Concentration: 1 mg/mL
• Dilution range: WB: 1 μg/mL; IHC: 2 μg/mL; IF: 20 μg/mL.Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human, mouse, and rat samples; Immunofluorescence in human and mouse samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 52 kDObserved: 60 kD
• Target: BECN1
• UniProt ID: Q14457
• NCBI: AAH10276
• Storage: Beclin-1 antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: Beclin-1 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: Beclin-1 Antibody: ATG6, VPS30, beclin1, GT197, Be
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

WB Validation in Human Cell Lines. Loading: 15 µg of lysate Antibodies: Beclin-1 orb1239257, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

WB Validation in Mouse Cell Lines. Loading: 15 µg of lysate Antibodies: Beclin-1 orb1239257, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

WB Validation in Rat Cell Lines. Loading: 15 µg of lysate Antibodies: Beclin-1 orb1239257, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Immunofluorescence Validation of Beclin-1 in HELA Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed Hela cells labeling Beclin-1 with orb1239257 at 10 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunofluorescence Validation of Beclin-1 in Mouse Brain Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain cells labeling Beclin-1 with orb1239257 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunohistochemistry Validation of Beclin-1 in Human Brain Tissue. Immunohistochemical analysis of paraffin-embedded human brain tissue using anti-Beclin-1 antibody (orb1239257) at 2.5 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of Beclin-1 in Mouse Kidney Tissue. Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-Beclin-1 antibody (orb1239257) at 1 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of Beclin-1 in Rat Liver Tissue. Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-Beclin-1 antibody (orb1239257) at 1 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of Beclin-1 in Rat Spleen Tissue. Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-Beclin-1 antibody (orb1239257) at 1 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Knockdown of Beclin-1 in Human T47D Cells (Thomas et al., 2011). T47D cells were transfected with scrambled or Beclin-1 siRNA. The following day, these cells were treated with vehicle, C, 0.5 mM VPA, 10 lM 4OH-tamoxifen, T, or 0.5 mM VPA and 10 lM 4OH-tamoxifen, VT, for 48 h and viability was assayed by dye exclusion.

Immunofluorescent Validation of Beclin-1 in Human Cortical Neurons and Rat Brain (Wang et al., 2017). Cell specificity of Beclin 1 expression. Cortical neurons and glia co-cultures (upper panels) and cortex slices (lower panels) were immunostainedwith anti-Beclin 1 (red) and NeuN (neuronal biomarker, green) orGFAP (astrocyte biomarker, green). Neurons and astrocytein cultured cells were indicated by arrows and arrowhead, respectively. Areas in white boxes were enlarged.

Immunohistochemistry Validation of Beclin-1 in Human HCC Cells (Qiu et al., 2014). Beclin-1 exhibited cytoplasmic staining in adjacent non-tumor (ANT) tissues (A), bordeling site between HCC and ANT (B: left, ANT; right: HCC), and HCC (C), respectively. Beclin-1 expression was stronger in ANT than in HCC. Magnification × 400.

Regulated Expression of Beclin-1 in Human T47D Cells (Thomas et al., 2011). Treatment with tamoxifen and an HDAC inhibitor alters the balance of autophagy and apoptosis inhibitors and drivers. T47D cells were treated with vehicle, C, 0.5 mM VPA, V, 10 lM 4OHtamoxifen, T, or 0.5 mM VPA and 10 lM 4OH-tamoxifen, VT, for 48 h and Beclin-1 protein and RNA.