BECN1 Recombinant Monoclonal Antibody
Description
Introduction to BECN1 and the Recombinant Monoclonal Antibody
BECN1 (Beclin-1) is a core autophagy-related protein critical for initiating autophagosome formation and regulating membrane trafficking pathways. It interacts with PI3K complexes (PI3KC3-C1 and PI3KC3-C2) to mediate phosphatidylinositol 3-phosphate production, essential for autophagy and endocytosis . Dysregulation of BECN1 is implicated in cancer, neurodegenerative diseases, and viral defense .
BECN1 Recombinant Monoclonal Antibody is a synthetic antibody engineered via in vitro systems to target BECN1 with high specificity. It is widely used in research to study BECN1’s role in autophagy, apoptosis, and disease mechanisms .
Applications and Reactivity
The antibody enables diverse experimental approaches:
Autophagy Regulation
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Autophagosome Formation: BECN1 knockout leads to malformed autophagosomes and impaired lipidation of LC3B-II, a marker of autophagosomes .
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PI3KC3 Complex Activation: BECN1 is required for PI3KC3 activity and omegasome formation, critical for autophagy initiation .
Ferroptosis and Disease
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Ferroptosis Induction: BECN1 phosphorylation (e.g., AMPK-mediated S93/96) promotes lipid peroxidation and ferroptosis by forming complexes with SLC7A11, inhibiting glutathione synthesis .
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Cancer and Neurodegeneration: Reduced BECN1 expression correlates with tumor progression in lung cancers , while dysregulation is linked to neurodegenerative diseases .
Subcellular Localization
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Mitochondrial Apoptosis: A 35 kDa truncated BECN1 isoform induces mitochondrial translocation of BAX and proapoptotic factor release .
Western Blot Challenges
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Band Discrepancy: Observed BECN1 bands often appear at ~60 kDa (vs. expected 52 kDa), likely due to post-translational modifications or alternative splicing .
Disease Relevance and Therapeutic Potential
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Cancer: BECN1 overexpression enhances chemosensitivity and ferroptosis in solid tumors, offering a therapeutic target .
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Neurodegeneration: BECN1’s role in clearing aggregates (e.g., amyloid-β) highlights its potential in treating Alzheimer’s disease .
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Viral Defense: BECN1 protects against Sindbis virus by promoting antiviral autophagy .
Product Specs
The BECN1 recombinant monoclonal antibody is produced using in vitro expression systems. These systems are developed by cloning the DNA sequences of BECN1 antibodies obtained from immunoreactive rabbits. The immunogen used is a synthesized peptide derived from the human BECN1 protein. Subsequently, the genes encoding the BECN1 antibodies are inserted into plasmid vectors, and these recombinant plasmid vectors are transfected into host cells for antibody expression. Following expression, the BECN1 recombinant monoclonal antibody undergoes affinity-chromatography purification. Rigorous testing in ELISA and FC applications confirms its reactivity with the human BECN1 protein.
BECN1 is a crucial component of the autophagic machinery and is essential for the initiation and regulation of autophagy, a cellular process playing a pivotal role in cellular quality control, homeostasis, and responses to stress. Dysregulation of BECN1 and autophagy has been implicated in various diseases, including cancer, neurodegenerative disorders, and infectious diseases.
Target Background
- A study showed that the expression of Beclin 1 was upregulated in 4-nitroquinoline 1-oxide-induced oral carcinogenesis, accompanied by myeloid-derived suppressor cells and regulatory T cells accumulation. PMID: 30272335
- Decreased Beclin-1 expression might be related to poor differentiation and unfavorable outcome in hepatocellular carcinoma. PMID: 30107804
- BECN1 knockout inhibited the migratory and invasive ability of MDAMB231 cells by partially reversing signals of epithelial-mesenchymal transition. Analysis of publicly available gene expression datasets revealed increased expression of BECN1 in triple-negative breast cancer samples. PMID: 30015871
- Low beclin 1 expression is associated with synovial sarcoma. PMID: 30066884
- ISG15 reduces miR-17-5p levels and thereby promotes Beclin-1-mediated autophagy. PMID: 29518394
- CaMKII can directly phosphorylate Beclin 1 at Ser90 to promote K63-linked ubiquitination of Beclin 1 and activation of autophagy; it also promotes K63-linked ubiquitination of inhibitor of differentiation 1/2 (Id-1/2) by catalyzing phosphorylation of Id proteins and recruiting TRAF-6. PMID: 29079782
- PFN1 could promote autophagy through participation in the Beclin1 complex and contribute to bortezomib resistance, which may become a novel molecular target in the therapy of MM. PMID: 29945297
- The results suggest that autophagy-associated proteins LC3A, LC3B, and Beclin-1 might be potential biomarkers for subclassification, differentiation, and local metastasis in primary lung tumor. PMID: 29545906
- Silencing Beclin-1 can attenuate hypoxia-induced VM formation and the metastatic ability of U87MG cells and is a potential target for VM inhibition in glioma. PMID: 29278874
- Endogenous HER2 interacts with Beclin 1 in breast cancer cells and inhibits autophagy. BECN1 allelic loss is common and associated with a worse prognosis in patients with HER2-overexpressing breast tumors. PMID: 29610308
- Phosphorylated and activated deoxycytidine kinase inhibits ionizing radiation (IR)-induced total cell death and apoptosis, and promotes IR-induced autophagy through the mTOR pathway and by inhibiting the binding of Bcl2 protein to BECN1 in breast cancer cells. PMID: 29393406
- Variation found in the BECN1 promoter modulates expression levels of this gene. Machado-Joseph Disease patients carrying the rs60221525 C allele presented a tendency for earlier estimated age at onset. Moreover, patients with the rs60221525 C allele presented a more severe clinical picture, compared to patients without this allele. PMID: 28699106
- that after Caspase 3-mediated Beclin 1 cleavage at 146/149 sites, the N-terminal fragments of Beclin 1 may partially translocate into nuclei in neurons subjected to AD-like injury. PMID: 28964771
- TRIM50, via the RING domain, ubiquitinates Beclin 1 in a K63-dependent manner enhancing its binding with ULK1 and autophagy activity. PMID: 29604308
- The expression level of EMC-6 is significantly elevated in cervical cancer, without significant correlation with Beclin1 and Rab5a. PMID: 28742203
- CISD2 was increased in glioma samples and was associated with poor prognosis and aggressive tumor behavior. The miR449a/CISD2/beclin1-mediated autophagy regulatory network contributed to the proliferation of glioma cells. PMID: 28983596
- Low expression levels of Beclin 1 may contribute towards the degeneration of chondrocytes. Beclin 1 overexpression increased cell viability, inhibited apoptosis and MMPs, likely via the PI3K/Akt/mTOR signaling pathway. PMID: 28731147
- A construct lacking the C-terminal BARA domain but including the coiled-coil region exhibits a homodimer KD 3.5-fold weaker than that of full-length BECN1, indicating that both the BARA domain and the coiled-coil region of BECN1 contribute to dimer formation. PMID: 29185708
- While 3-methyladenine rescues cell damage. Our data thus suggest that I/R promotes NF-kappaB p65 activity mediated Beclin 1-mediated autophagic flux, thereby exacerbating myocardial injury. PMID: 27857190
- that TPT1 knockdown altered the BECN1 interactome, a representative MTOR-independent pathway, to stimulate autophagosome formation. PMID: 28409693
- High levels of Beclin1 is associated with non-small cell lung cancer. PMID: 28558758
- OHPg/PR-B through Beclin-1 and Bcl2 evoke autophagy-senescence transition in breast cancer cells. PMID: 27462784
- These results suggest that the phosphorylated CAV1 functions to activate autophagy through binding to the BECN1/VPS34 complex under oxidative stress and to protect against ischemic damage. PMID: 28542134
- some tumor suppressor functions of Beclin 1 were mediated through posttranscriptional regulation of ZEB1 via AUF1 in thyroid cancers. PMID: 27683118
- Cheliensisin A (Chel A) treatment led to PH domain and Leucine rich repeat Protein Phosphatases (PHLPP2) protein degradation and subsequently increased in c-Jun phosphorylation, which could be attenuated by inhibition of autophagy mediated by Beclin 1. PMID: 27556506
- autophagy induction stabilized microtubules by degrading SCG10, a microtubule disassembly protein in neurons. In mice with spinal cord injury, local administration of a specific autophagy-inducing peptide, Tat-beclin1, to lesion sites markedly attenuated axonal retraction of spinal dorsal column axons and cortical spinal tract and promoted regeneration of descending axons following long-term observation. PMID: 27638205
- Results provide evidence that Beclin-1 plays a major role in breast cancer subtypes, and identified AFF3 as his direct target. PMID: 28401970
- significantly different gene expressions of BECN1 and PRKCB between the control and the Alzheimer's disease (AD) groups and of CDKN2A between the control and the preclinical AD groups, are reported. PMID: 26510741
- EHMT2 can directly repress Beclin-1 and the inhibition of EHMT2 may be a useful therapeutic approach for cancer prevention by activating autophagy. PMID: 27174920
- oxidative stress activates the TRPM2-Ca(2+)-CAMK2 cascade to phosphorylate BECN1 resulting in autophagy inhibition. PMID: 27245989
- Inhibition of Beclin-1 resulted in a dramatic decline of autophagy and increase of apoptosis in drug-resistant cells, accompanied by a remarkable reduction in S phase and a raise in G2/M phase of cell cycle and reverse the effect induced by miR-30a-5p suppression. PMID: 28977798
- AMP-activated protein kinase (AMPK) regulates autophagy by phosphorylating BECN1 at Thr388. PMID: 27304906
- In patients treated with immunomodulatory drugs (IMiDs) lack of or low Beclin-1 expression resulted in a significantly improved OS and PFS compared to those treated with bortezomib or nonnovel agents. PMID: 26880040
- beta-asarone changed the cellular morphology, inhibited cell proliferation, and enhanced the expression of P53, LC3-II/I, Beclin-1, AMPK, and pAMPK while inhibiting the expression of P62, Bcl-2, mTOR, and pmTOR in glioma cells. PMID: 28776671
- PINK1 silencing impaired BECN1 enrichment at mitochondria-associated membranes independently of PARK2, suggesting a novel role for PINK1 in regulating mitophagy. PMID: 28368777
- ABHD5 possesses a PNPLA2-independent function in regulating autophagy and tumorigenesis. PMID: 27559856
- BECN1-APP association and BECN1-dependent APP endocytosis and degradative trafficking were negatively regulated by active AKT. PMID: 27715386
- findings will also clarify the conflict between apoptosis-autophagy on the basis of its unique property derived from surface chemistry modulation, in context of Beclin-1 eminent cleavage/splice which remarks novel effect of Beclin-1 dependent tuning of autophagosomes formation and switch to enhance apoptotic index, mediates by PI3KC3 cleavage and caspase activation. PMID: 28076888
- Data suggest that BECN1 exhibits a helical region that transitions between packing (folding/unfolding) as part of either one of two conserved domains; packing of this helical region appears to have important implications for relative stability of autophagy-inactive and autophagy-active BECN1 complexes. PMID: 28798234
- downregulation of HOXC9 releases its transcriptional inhibition of DAPK1, resulting in the activation of the DAPK1-Beclin1 pathway, which induces autophagy in glioblastoma cells. PMID: 26582930
- ASPP2 is a key regulator of BECN1-dependent autophagy. PMID: 27929538
- the polyQ domain enables wild-type ataxin 3 to interact with beclin 1, a key initiator of autophagy; this interaction allows the deubiquitinase activity of ataxin 3 to protect beclin 1 from proteasome-mediated degradation and thereby enables autophagy. PMID: 28445460
- findings suggest that expression of cFLIPL regulates the basal interaction of Bcl-2 with Beclin-1 and substantiates p53 dependent ubiquitination of Beclin-1 during autophagic stress to determine the fate of cell death or survival. PMID: 26682748
- Here, sequence analyses, structural modeling, mutagenesis combined with pull-down assays, X-ray crystal structure determination and small-angle X-ray scattering were used to investigate the Beclin 1-GAPR-1 interaction. changes in the structure of the equatorial groove combined with the improved dimerization of pentad mutant GAPR-1 are likely to abrogate binding to Beclin 1. PMID: 28876241
- Acetylated PGK1 binds to and phosphorylates Beclin1 at S30, leading to activation of the VPS34-Beclin1 complex to initiate autophagosomal formation. PMID: 28238651
- reactive oxygen species (ROS) production was induced, which increased RelA transcriptional activity and beclin 1 (BECN1) expression in oral cancer cells irradiated with LPL. PMID: 27632526
- Further data showed that reactive oxygen species-c-Jun activation by HBx resulted in the release of beclin-1 from its association with beclin-2 to form a complex with VPS34, thus enhancing autophagosome formation. PMID: 28515304
- Beclin1 interacts with both the N- and C-termini of LAPTM4B. Beclin1 competes with EGFR for LAPTM4B binding. PMID: 28479384
- MiR-30d inhibited cell autophagy by directly targeting messenger RNA of ATG5, phosphoinositide 3-kinase, and Beclin1 and promoted cell apoptosis of human colon cancer cells. It is helpful to clarify the function of miR-30d in tumorigenesis of human cancers. PMID: 28651493
- fructose-1,6-bisphosphatase 1 facilitated co-action between Bcl-2 and Beclin 1, which may be important in the mechanism of fructose-1,6-bisphosphatase 1-mediated mitophagy inhibition. In summary, loss of mitophagy by fructose-1,6-bisphosphatase 1-mediated repression promotes apoptosis in breast cancer. PMID: 28653874
Q&A
What species reactivity can be expected from BECN1 monoclonal antibodies?
Most commercially available BECN1 recombinant monoclonal antibodies demonstrate cross-reactivity across multiple species. These typically include human, mouse, and rat samples, with some antibodies showing extended reactivity to monkey models . When selecting an antibody for your research, confirm the specific cross-reactivity profile, as it varies by manufacturer. For example, the human/mouse/rat Beclin 1 antibody MAB5295 shows validated detection in human epithelial carcinoma (A431), mouse B cell lymphoma (A20), and rat normal kidney (NRK) cell lines . Some antibodies may also have predicted reactivity to additional species such as pig, zebrafish, bovine, and others, though these require experimental validation .
What applications are BECN1 recombinant monoclonal antibodies validated for?
BECN1 recombinant monoclonal antibodies are validated for multiple research applications:
When designing experiments, optimal dilutions should be determined empirically for your specific application and sample type .
How is BECN1 recombinant monoclonal antibody produced compared to conventional monoclonal antibodies?
BECN1 recombinant monoclonal antibodies are created through in vitro expression systems rather than traditional hybridoma technology. The production process involves:
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Cloning DNA sequences from immunoreactive animals (typically rabbits)
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Using a synthesized peptide derived from human BECN1 protein as the immunogen
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Inserting genes encoding BECN1 antibodies into plasmid vectors
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Transfecting these recombinant plasmid vectors into host cells
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Expressing the antibody in the host cell system
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Purifying the expressed antibody through affinity-chromatography
This recombinant approach provides superior lot-to-lot consistency, continuous supply, and animal-free manufacturing compared to conventional methods . The antibodies undergo rigorous testing in applications such as ELISA and flow cytometry to confirm reactivity with the human BECN1 protein .
How can I optimize detection of BECN1 in Western blot applications?
For optimal detection of BECN1 in Western blot applications, consider the following protocol optimizations:
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Sample preparation: Prepare lysates from appropriate cells with confirmed BECN1 expression (e.g., A431 human epithelial carcinoma, A20 mouse B cell lymphoma, or NRK rat normal kidney cell lines) .
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Membrane selection: PVDF membranes show good results for BECN1 detection .
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Antibody concentration: Start with 0.1-1 μg/mL of anti-BECN1 antibody, followed by an appropriate HRP-conjugated secondary antibody .
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Reducing conditions: Perform the experiment under reducing conditions for optimal band detection .
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Buffer system: Use appropriate buffer systems such as Immunoblot Buffer Group 2 for consistent results .
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Expected band size: Look for a specific band at approximately 60 kDa, though this may vary slightly (58-60 kDa) depending on sample type and experimental conditions .
For troubleshooting, positive controls using lysates from HeLa cells, HEK-293 cells, HepG2 cells, or MCF-7 cells can confirm antibody functionality, as these have been validated in multiple antibody systems .
What are the considerations for studying BECN1 interactions with other proteins?
When studying BECN1 interactions with other proteins, such as APP (amyloid β precursor protein), consider these methodological approaches:
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Co-immunoprecipitation (Co-IP):
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Surface protein interaction studies:
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Controls:
This approach revealed that BECN1 interacts with APP at the cell surface, suggesting a role in regulating the endocytosis of surface APP and sorting it for lysosomal degradation .
How can BECN1 knockdown be validated when studying its functional role?
When validating BECN1 knockdown for functional studies:
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shRNA approach:
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Confirmation methods:
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Functional validation:
What considerations are important for immunohistochemical detection of BECN1 in tissue samples?
For successful immunohistochemical detection of BECN1 in tissue samples:
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Tissue preparation:
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Epitope retrieval:
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Antibody concentration and incubation:
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Detection and counterstaining:
How can I design experiments to study BECN1's role in autophagy regulation?
To investigate BECN1's role in autophagy regulation:
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Protein-protein interaction studies:
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Cell trafficking experiments:
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Study BECN1's role in trafficking of autophagy-related proteins
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Use the cell surface biotinylation approach to track internalization rates
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Label surface proteins at 4°C, then facilitate internalization at 37°C for defined time periods (e.g., 5 minutes)
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Compare internalization rates between wild-type and BECN1-depleted cells
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Quantification methods:
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For internalization studies, analyze both the total amount of protein on the cell surface before internalization and the internalized pool after acid treatment (to remove residual surface antibodies)
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Use immunofluorescence microscopy to visualize differences between control and BECN1-manipulated cells
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Quantify results through appropriate image analysis software
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Controls and validation:
