Antibodies against beclin 1

Manufactured by Cell Signaling Technology

Sourced in United States

Antibodies against beclin-1 are laboratory reagents used to detect and analyze the beclin-1 protein. Beclin-1 is a key regulator of autophagy, a cellular process involving the degradation and recycling of cellular components. These antibodies can be used in various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression, localization, and function of beclin-1 in biological systems.

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3 protocols using antibodies against beclin 1

Western Blot Analysis of Protein Markers

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For Western blot analyses, cells were scraped, and lysed in buffer A containing 10 mM HEPES, 10 mM NaCl, 1.5 mM MgCl2, 0.25% Tween 20, 1mM dithiothreitol, 100 mg/ml phenylmethylsulfonyl fluoride, 1 mg/ml leupeptin, and 15 mg/ml aprotinin. Thirty μg of total protein from each sample was then loaded onto 10~15% SDS-polyacrylamide gels, because all sorts of proteins measured in this experiment have a diversity of molecular weights (from 4 ~ 120 kDa). Separated proteins were transferred to nitrocellulose membranes (GE Healthcare Life Sciences, Piscataway, NJ, USA), which were blocked with 5% skim milk (at 4°C overnight), and incubated with antibodies against SIRT1 (Santa Cruz, CA), anti-Aβ (6E10) (Covance, Emeryville, CA), and rabbit polyclonal CTFβ (751–770) (Calbiochem, La Jolla CA. Antibodies against beclin-1, Atg5, and LC3 (rabbit monoclonal LC3A/B, 1:1000) were obtained from Cell Signaling Technology (Danvers, MA). Membranes were reprobed with an anti-β-actin antibody (MP Biomedicals, LLC, Aurora, OH, USA) as an internal control.

Lee H.R., Shin H.K., Park S.Y., Kim H.Y., Bae S.S., Lee W.S., Rhim B.Y., Hong K.W, & Kim C.D. (2015). Cilostazol Upregulates Autophagy via SIRT1 Activation: Reducing Amyloid-β Peptide and APP-CTFβ Levels in Neuronal Cells. PLoS ONE, 10(8), e0134486.

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Glioblastoma Therapeutic Evaluation Protocol

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2-BP (18,263-25-7) was purchased from Merck (Darmstadt, Germany). TMZ (HY-17364) was purchased from MedChemExpress (New Jersey, USA). Antibodies against P62 (1:1,000; ab109012), LC3B (1:1,000; ab192890), and Bcl2 (1:1,000; ab196495) were purchased from Abcam (Cambridgeshire, England). Antibodies against Beclin-1 (1:1,000; no. 3495), PCNA (1:1,000; no. 13110), Bcl-xl (1:1,000; no. 2764), and caspase-3 (1:1,000; no. 9662) were purchased from Cell Signaling Technology (Massachusetts, USA). Antibodies against GAPDH (1:1,000; bs-0755R) were purchased from Bioss (Beijing, China). A goat anti-rabbit secondary antibody (1:10,000; lot: 70,100,200) was purchased from Biosharp (Wuhan, China).

Tang F., Yang C., Li F.P., Yu D.H., Pan Z.Y., Wang Z.F, & Li Z.Q. (2022). Palmitoyl transferases act as potential regulators of tumor-infiltrating immune cells and glioma progression. Molecular Therapy. Nucleic Acids, 28, 716-731.

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Regulation of STAT3-Dependent Autophagy

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STAT3-dependent luciferase reporter plasmid was provided by Dr Ming Shi from our institute. Human HO-1, STAT3, ATG5 and Src siRNA and their control siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), Invitrogen-Life Technology (Beijing, China) and Ruibo Biotechnology (Guangzhou, China), respectively. The antibodies against Beclin-1, LC3B, phospho-Tyr416-Src, Src, phospho-Tyr705-STAT3 and STAT3 were purchased from Cell Signaling Technology (Beverly, MA, USA). The antibodies against HO-1 and β-actin were obtained from Santa Cruz Biotechnology. The anti-ATG5 antibody, DOX, 3-Methyladenine (3-MA) and chloroquine were purchased from Sigma (St. Louis, MO, USA).

Tan Q., Wang H., Hu Y., Hu M., Li X., A., Ma Y., Wei C, & Song L. (2015). Src/STAT3-dependent heme oxygenase-1 induction mediates chemoresistance of breast cancer cells to doxorubicin by promoting autophagy. Cancer Science, 106(8), 1023-1032.

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