Metformin (D150959), CQ (C6628), MDC (30432), 3-MA (M9281), 4',6-diamidino-2- phenylindole (DAPI; D9542), antibodies against anti-β and LC3, anti-rabbit secondary antibody, anti-mouse secondary antibody and Cy3-labeled rabbit secondary antibody were purchased from Sigma (St. Louis, MO, USA). Antibodies against p-Stat3 (Y705), Stat3, cyclin D1, LC3, PCNA and PARP were purchased from Cell Signaling (Beverly, MA, USA); antibodies against p-AMPK, p-mTOR, Beclin-1, Bcl-2, p62 and Bax were purchased from Santa Cruz (Santa Cruz, CA, USA). Anti-phospho-Histone H3 (Ser10) antibody was purchased from Upstate (Millipore, Billerica, MA, USA). Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum were obtained from Gibco (Life Technologies Gibco/BRL, New York, NY, USA). Crystal violet staining and JC-1 were obtained from Beyotime Institute of Biotechnology (Shanghai, China). ApopTag plus peroxidase in situ apoptosis detection kit was obtained from Millipore. AO was obtained from Poly-Sciences (Warrington, PA, USA). Stat3, AMPK, Beclin-1 and Atg5 siRNAs were purchased from Shanghai GenePharma (Shanghai, China). Lipofectamine 2000 was bought from Invitrogen (Carlsbad, CA, USA).
Beclin1
Manufactured by GenePharma
Sourced in China
Beclin1 is a protein involved in the regulation of autophagy, a cellular process that degrades and recycles damaged or unwanted cellular components. Beclin1 plays a crucial role in the initiation and formation of autophagosomes, which are the double-membrane vesicles that engulf the targeted cellular material for degradation. This protein is essential for the proper functioning of the autophagic pathway and has been studied extensively in the context of various biological processes and disease conditions.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Control sirna, by GenePharma (3 mentions)
Stat3, by GenePharma (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Crystal violet staining, by Beyotime (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Cy3 labeled rabbit secondary antibody, by Merck Group (1 mentions)
Atg5 sirna, by GenePharma (1 mentions)
Anti rabbit secondary antibody, by Merck Group (1 mentions)
Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (1 mentions)
Beclin 1, by Santa Cruz Biotechnology (1 mentions)
Apoptag plus peroxidase in situ apoptosis detection kit, by Merck Group (1 mentions)
Bcl2 associated x (bax), by Santa Cruz Biotechnology (1 mentions)
Bcl 2, by Santa Cruz Biotechnology (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Beclin 1, by Cell Signaling Technology (1 mentions)
Chloroquine diphosphate cq, by Selleck Chemicals (1 mentions)
Jujuboside b, by Merck Group (1 mentions)
10 protocols using beclin1
1
Autophagy Modulation in Cancer Therapeutics
2
Transfection of Autophagy Regulators
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The siRNAs to Atg5, Beclin-1, Stat3, AMPK or control siRNA were all purchased from Shanghai GenePharma. Cells were transfected with siRNA using Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions. Cells were incubated for 48 h before further treatment.
3
Beclin 1 knockdown in luteal cells
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To knockdown the Beclin 1 expression, the siRNAs-targeting Beclin1 and nontargeting scramble siRNA were synthesized by GenePharma, China. The sequences of siRNAs were as follows: Beclin 1 siRNA1, sense, 5′-CUC AGG AGA GGA GCC AUU UTT-3′, antisense, 5′-AAA UGG CUC CUC UCC UGA GTT-3′ and negative control siRNA, sense, 5′-UUC UCC GAA CGU GUC ACG UTT-3′, antisense, 5′-ACG UGA CAC GUU CGG AGA ATT-3′. Luteal cells were seeded at 2 × 105 per well in 6-well plates overnight, and the transfection was performed, when the cells reached a 70% confluence. During transfection, siRNA duplexes (50 pmol) were transfected into the target cell populations using the Lipofectamine 3000 Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. Culture media were collected for progesterone measurements 48 h after transfection, and cell proteins were extracted using RIPA for assessing protein expression levels by western blotting.
4
Jujuboside B Modulates Autophagy Pathways
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Jujuboside B was purchased from Sigma and dissolved in dimethyl sulfoxide (DMSO) to the concentration of 100 mM stock solution kept at −20°C. The primary antibodies to cleaved PARP, total PARP, LC3, p62, NOXA, beclin1, ATG7, AMPK, and phospho-AMPK were all purchased from Cell Signaling Technology (United States). The primary antibodies to β-actin and GAPDH were purchased from Santa Cruz Biotechnology (CA, United States). Antibodies to cleaved caspase-3, total caspase-3, and the secondary HRP-conjugated anti-rabbit and anti-mouse antibodies were purchased from HUABIO. BCA protein assay kit and protein page ruler were purchased from Thermo Fisher. The knockdown siRNA for NOXA, beclin1, ATG7, AMPK, and control siRNA were purchased from GenePharma (Shanghai, China). The autophagy inhibitor chloroquine diphosphate (CQ) was purchased from Selleck.
5
Modulating HDAC9, β-catenin, and Beclin1 in Hypoxic Myoblasts
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siRNA duplex oligonucleotides against mouse HDAC9 (Gene-Pharma Co, Shanghai, China), β-catenin (Gene-Pharma Co, Shanghai, China), Beclin1 (Gene-Pharma Co), or the negative control (Gene-Pharma Co) were transfected into both normoxic and hypoxic C2C12 myoblasts at a final concentration of 50 nM using siPORTNeoFX. The medium was replaced 8 h later. Experiments were performed in triplicate.
6
Rat Cortical Neuron Transfection Protocols
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The rat cortical neurons (RN-c, MZ-7885) were purchased from Mingzhou Biotechnology (Ningbo, China), and the RN-c cell complete culture medium was from ORiCells Biotechnology (Shanghai, China).
Cell transfection was conducted based on the instructions of riboFECTTMCP transfection kit produced by RiboBio (Guangzhou, China), followed by 6-h incubation before subsequent experiments. Beclin-1 overexpression and interference plasmids (oe-Beclin-1 and sh-Beclin-1), METTL3 overexpression and interference plasmids (oe-METTL3 and sh-METTL3), KAT2A overexpression and interference plasmids (oe-KAT2A and sh-KAT2A), NRF1 interference plasmid (sh-NRF1), and their negative controls (oe-NCs and sh-NCs) were obtained from GenePharma (Shanghai, China).
Cell transfection was conducted based on the instructions of riboFECTTMCP transfection kit produced by RiboBio (Guangzhou, China), followed by 6-h incubation before subsequent experiments. Beclin-1 overexpression and interference plasmids (oe-Beclin-1 and sh-Beclin-1), METTL3 overexpression and interference plasmids (oe-METTL3 and sh-METTL3), KAT2A overexpression and interference plasmids (oe-KAT2A and sh-KAT2A), NRF1 interference plasmid (sh-NRF1), and their negative controls (oe-NCs and sh-NCs) were obtained from GenePharma (Shanghai, China).
7
Modulating Autophagy Regulators in HEK293T Cells
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siRNA targeting Atg5, Beclin1, and p62 were purchased from GenePharma (Shanghai, China) and were transfected into HEK293T cells using Lipofectamine 2000 (Invitrogen) with Opti-MEM® I Reduced Serum Medium (Gibco) according to the manufacturer's instructions. The siRNA sequences could be found in . pcDNA3-Flag-FOXO1 and GFP-FOXO3 constructs were kind gifts from Dr. Kun-Liang Guan (University of California, San Diego) and Dr. Mien-Chie Hung (MD Anderson Cancer Center), respectively, and have been previously described [44 (no link found, link), 45 (link)]. The plasmids were transfected into HEK293T cells with empty vectors as the negative control using X-tremeGENE HP DNA Transfection Reagent (Roche) according to the manufacturer's instructions.
8
Silencing Autophagy Genes in HCT116 Cells
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Atg5, Beclin 1, 5-LOX and negative control siRNA were synthesized by Genepharma. The sequences of siRNA were as following: human Atg5 siRNA, sense 5′-GAC GUU GGU AAC UGA CAA ATT-3′ and antisense 5′-UUU GUC AGU UAC CAA CGU CTT-3′; human Beclin 1siRNA, sense 5′-GGA GCC AUU UAU UGA AAC UTT-3′ and antisense 5′-AGU UUC AAU AAA UGG CUC CTT-3′. 5-LOX was designed according to previous study (targeting sequence:5′-GCGCAAGTACTGGCTGAATGA-3′ ; NM_000698) [52] (link). The siRNA were transfected with Lipofectamine 2000 reagent (Invitrogen, 11668027) for 24 h in HCT116 cells according to the manufacturer’s protocol.
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Atg5, Beclin 1, 5-LOX and negative control siRNA were synthesized by Genepharma. The sequences of siRNA were as following: human Atg5 siRNA, sense 5′-GAC GUU GGU AAC UGA CAA ATT-3′ and antisense 5′-UUU GUC AGU UAC CAA CGU CTT-3′; human Beclin 1siRNA, sense 5′-GGA GCC AUU UAU UGA AAC UTT-3′ and antisense 5′-AGU UUC AAU AAA UGG CUC CTT-3′. 5-LOX was designed according to previous study (targeting sequence:
9
Knockdown and Knockout of PCDRlnc1 in PC3-DR Cells
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Transfections were performed using Lipofectamine 2000 reagent (Invitrogen, USA) according to the manufacturer's instructions. The vector pcDNA3.1-PCDRlnc1 was purchased from GenePharma. Beclin-1 and UHRF-1 siRNAs (GenePharma, China) were used to knock down their gene expression. The sense sequences of siRNAs were as follows: Beclin-1: 5'-CAGTTTGGCACAATCAATA-3'; URHF1: 5'-GCGCUGGCUCUCAACUGCU-3'. The sequence of non-targeting control siRNA (si-NC) was 5'-UUCUCCGAACGUGUCACGUTT-3'. The efficiency was assessed after 48h using qRT-PCR or WB. CRISPR-Cas9 for PCDRlnc1 knockout CRISPR/CAS gene-editing technology mediated by electroporation was used to knockout PCDRlnc1 in PC3-DR cells (Cyagen, China). The loss of PCDRlnc1 in PC3-DR cells was verified by PCR and sequencing Figure S1 and Figure S2 .
10
Beclin1, TFEB Knockdown Protocol
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The siRNAs to Beclin1, TFEB or control siRNA were all purchased from Shanghai GenePharma. Cells were transfected with siRNA using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer's instructions. Cells were incubated for 48 h before further treatment.
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