BECN1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift: 93.41%.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift: 93.41%
Alternative names
APG6, ATG 6, ATG6 autophagy related 6 homolog, BECN1_HUMAN, Bcl-2-interacting protein beclin, Beclin 1 (coiled coil moesin like BCL2 interacting protein), Beclin 1 autophagy related, Beclin-1, Coiled-coil myosin-like BCL2-interacting protein, GT197, Protein GT197, VPS 30
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BECN1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift: 93.41%.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift: 93.41%
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- BECN1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM (High Glucose) + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Beclin 1 sometimes referred to as beclin-1 or becn1 is a protein that plays a mechanical role in autophagy. This process involves the recycling of cellular components and is essential for maintaining cellular health. Beclin 1 is approximately 60 kDa in molecular weight and is expressed in a variety of tissues throughout the body. It acts at the molecular level engaging in the initial stages of autophagosome formation which are critical for cell survival during nutrient deprivation.
- Biological function summary
The protein functions as an important component of the Beclin 1 autophagy complex which orchestrates the autophagic process. This complex is essential for promoting autophagosome nucleation by interacting with multiple partners that regulate the autophagy pathway. In addition to its role in autophagy Beclin 1 protein stabilizes interactions with other autophagic proteins ensuring the correct assembly and activity of this cellular mechanism. The regulation of Beclin 1's activity and expression influences the efficiency and specificity of autophagy in cells.
- Pathways
Beclin 1 integrates into critical cellular pathways like the PI3K/Akt pathway and the mTOR pathway which are pivotal in cell growth and survival. Within these pathways beclin cooperates closely with proteins such as ATG14 and VPS34. These interactions facilitate the recruitment and activation of additional factors that promote the elongation of phagophores which eventually become complete autophagosomes. The coordination of these pathways ensures the proper balance between cell survival and death adapting to cellular conditions.
- Associated diseases and disorders
Beclin 1 shows significant relevance in cancer and neurodegenerative diseases. Abnormal Beclin 1 expression and function are linked to a range of cancers as its dysregulation affects apoptosis and autophagic turnover. Furthermore in neurodegenerative disorders like Alzheimer's disease altered Beclin 1 activity can disrupt normal protein degradation exacerbating protein aggregation a hallmark of these conditions. Interactions with proteins like Bcl-2 are essential in the context of these diseases as Bcl-2 modulates the autophagic and apoptotic balance directly impacting disease progression.
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3 product images
Western blot - Human BECN1 (Beclin 1) knockout HeLa cell line (ab262511)
Lanes 1 - 2: Merged signal (red and green). Green - Anti-Beclin 1 antibody [OTI4A10] ab118148 observed at 60 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.Anti-Beclin 1 antibody [OTI4A10] ab118148 was shown to react with Beclin 1 in wild-type HeLa cells in western blot. Loss of signal was observed when BECN1 knockout sample was used. Wild-type HeLa and BECN1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in non-mammalian (TBS-based) blocking solution before incubation with Anti-Beclin 1 antibody [OTI4A10] ab118148 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Beclin 1 antibody [OTI4A10] (Anti-Beclin 1 antibody [OTI4A10] ab118148) at 1/1000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: BECN1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human BECN1 (Beclin 1) knockout HeLa cell line (ab262511)
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 60 kDa
Western blot - Human BECN1 (Beclin 1) knockout HeLa cell line (ab262511)
Western blot: Anti-BECN1 antibody [EPR19662] (Anti-Beclin 1 antibody [EPR19662] ab207612) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Beclin 1 antibody [EPR19662] ab207612 was shown to bind specifically to BECN1. A band was observed at 57 kDa in wild-type HeLa cell lysates with no signal observed at this size in BECN1 knockout cell line. To generate this image, wild-type and BECN1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Beclin 1 antibody [EPR19662] (Anti-Beclin 1 antibody [EPR19662] ab207612) at 1/2000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: BECN1 knockout HeLa cell lysate at 20 µg
Lane 3: Jurkat cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Next Generation Sequencing - Human BECN1 (Beclin 1) knockout HeLa cell line (ab262511)
4 bp deletion after Gln74 of the WT protein
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