Mouse anti beclin1

Manufactured by BD

Sourced in United States

Mouse-anti-Beclin1 is a laboratory reagent used for the detection and analysis of Beclin1 protein in biological samples. Beclin1 is a key regulator of autophagy, a cellular process involved in the degradation and recycling of damaged or unwanted cellular components. This product can be used in various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of Beclin1 in different cell types or tissues.

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3 protocols using mouse anti beclin1

Western Blot Analysis of Autophagy Proteins

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Cells in microslides were washed twice with ice-cold PBS and lysed in ice with 150 µl of 1X Laemmli buffer (60 mM Tris-HCL pH = 6.8, 2% SDS, 10% Glycerol, bromophenol blue, supplemented with 100 mM DTT) for 30 min. Samples were boiled for 10 min at 95 °C, separated by SDS/PAGE and then transferred onto Nitrocellulose membranes. Western blot analysis was performed with specific antibodies and the antigen–antibody complexes were visualized by chemiluminescence (Immobilon Western, Merck Millipore). The following antibodies were used in immunoblotting: rabbit-anti LC3 (1:1000, Sigma, Cat#L7543); mouse-anti-actin (1:5000, Millipore, Cat#1501); mouse-anti-Beclin1 (1:2000, BD Biosciences,Cat#612113); rabbit-anti-IFT88 (1:1000, Proteintech, Cat#13967-1-AP); rabbit-anti-PIK3C2α (1:800, Novus, Cat#NBP2-19829); mouse-anti-Wipi2 (2A2) (1:1000, AbD Serotec, Cat#MCA5780GA); rabbit-anti-Rab11 (1:1000, Cell Signaling, Cat#2413S); rabbit-anti-VPS34 (d9A5) (1:500, Cell Signaling, Cat#4263); rabbit-anti-LKB1 (1:1000, Cell signaling, Cat#3050); rabbit-anti-FIP200 (1:1000, Sigma, Cat#SAB4200135); mouse-anti-ATG16L1 (1:1000, MBL, Cat#PM040); rabbit-anti-IFT20 (1:1500, Proteintech, Cat#13615). Secondary HRP conjugate anti‐rabbit IgG (GE Healthcare) and HRP conjugate anti‐mouse IgG (Bio‐Rad).

Boukhalfa A., Nascimbeni A.C., Ramel D., Dupont N., Hirsch E., Gayral S., Laffargue M., Codogno P, & Morel E. (2020). PI3KC2α-dependent and VPS34-independent generation of PI3P controls primary cilium-mediated autophagy in response to shear stress. Nature Communications, 11, 294.

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Comprehensive Antibody Panel for Cellular Analysis

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The following primary antibodies were employed for either immunofluorescence or western blotting: mouse anti-ARH-I (for IF 1:250, for WB 1:1000; cod. ab45768; Abcam, Cambridge, UK), mouse anti-p21 (for IF 1:100, for WB 1:200; cod. sc-817; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-LC3 (1:2000; cod. L7543; Sigma-Aldrich), rabbit anti-p62 (1:500; cod. 8025; Cell Signaling, Danvers, MA, USA), mouse anti-β-actin (1:2000; cod. A5441; Sigma-Aldrich), mouse anti-BECLIN-1 (1:500; cod. 612112; BD Biosciences, Franklin Lakes, NJ, USA), goat anti-BECLIN-1 (1:250; cod. sc-10086; Santa Cruz Biotechnology), rabbit anti-VPS34 (1:500; cod. 4263; Cell signaling), mouse anti-STAT3 (1:500; cod. 9139; Cell Signaling), rabbit anti-phospho (Tyr705) STAT3 (1:1000; cod. 9145; Cell Signaling), rabbit anti-Ki-67 (1:100; cod. HPA001164; Sigma-Aldrich), mouse anti-BCL-2 (1:500; cod. 15071; Cell Signaling), mouse anti-IL-6R (1:1000; cod. AHR0061; Invitrogen, Waltham, MA, USA), rabbit anti-cyclin D1 (1:500; cod. 2978; Cell Signaling), rabbit anti-p38 (1:500; cod. 9212; Cell Signaling), rabbit anti-GAPDH (1:1000, cod. G9545, Sigma Aldrich), and mouse anti-ERK1/2 (1:500; cod. 05-1152; Millipore, Burlington, MA, USA).

Esposito A., Ferraresi A., Salwa A., Vidoni C., Dhanasekaran D.N, & Isidoro C. (2022). Resveratrol Contrasts IL-6 Pro-Growth Effects and Promotes Autophagy-Mediated Cancer Cell Dormancy in 3D Ovarian Cancer: Role of miR-1305 and of Its Target ARH-I. Cancers, 14(9), 2142.

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Molecular Cloning and Tagging Strategies

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Human Dpr1 cDNA was cloned into pEGFP-C3 to generate GFP-Dpr1. Human Vps34 cDNA was cloned into the KpnI-XbaI sites of the pcDNA3.1(+)-HA or pCS2(+)-Flag vector. The Flag-Beclin1 was a gift from Dr Honggang Wang. The p62-GFP was a gift from Dr Terje Johansen. Dvl2 was cloned into the Bam HI site of pDsRed-N1. Atg14L was cloned from Atg14L-GFP into EcoRI-SalI sites of pmCherry-N1. Dpr1 shRNAs were purchased from Open Biosystems. MG132 was from Calbiochem and prepared as 100 mg/ml in DMSO. BafA1, rapamycin, and XAV939 were obtained from Sigma. A769662 was from Abcam. Antibodies include: mouse anti-Calbindin (Sigma), rabbit anti-GFAP (Bioss, Beijing), mouse anti-ubiquitin (FK2 MBL), mouse anti-Beclin1 (BD Biosciences), rabbit anti-Beclin1 (MBL), rabbit anti-Vps34 (Cell Signaling), rabbit anti-Atg14L (MBL), mouse anti-Flag M2 (Sigma), rabbit anti-Dvl2 (Cell Signaling), mouse anti-tubulin and anti-GFP (Santa Cruz Biotech), mouse anti-p62 and rabbit anti-LC3 (MBL).

Ma B., Cao W., Li W., Gao C., Qi Z., Zhao Y., Du J., Xue H., Peng J., Wen J., Chen H., Ning Y., Huang L., Zhang H., Gao X., Yu L, & Chen Y.G. (2014). Dapper1 promotes autophagy by enhancing the Beclin1-Vps34-Atg14L complex formation. Cell Research, 24(8), 912-924.

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