Anti-Beclin 1 antibody [OTI4A10] (ab118148)
Key features and details
- Mouse monoclonal [OTI4A10] to Beclin 1
- Suitable for: WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-Beclin 1 antibody [OTI4A10]
See all Beclin 1 primary antibodies -
Description
Mouse monoclonal [OTI4A10] to Beclin 1 -
Host species
Mouse -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein corresponding to Human Beclin 1. Produced in HEK-293T cells. NP_003757
Database link: Q14457 -
Positive control
- WB: Beclin 1 transfected HEK-293T cell lysate; HeLa cell lysate.
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General notes
The clone number has been updated from 4A10 to OTI4A10, both clone numbers name the same clone.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.30
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA, 50% Glycerol -
Concentration information loading...
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Purity
Affinity purified -
Purification notes
Purified from cell culture supernatant by affinity chromatography -
Clonality
Monoclonal -
Clone number
OTI4A10 -
Isotype
IgG2a -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab118148 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/2000. Predicted molecular weight: 52 kDa.
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Notes |
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WB
1/2000. Predicted molecular weight: 52 kDa. |
Target
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Function
Plays a central role in autophagy (By similarity). May play a role in antiviral host defense. Protects against infection by a neurovirulent strain of Sindbis virus. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the beclin family. -
Cellular localization
Golgi apparatus > trans-Golgi network membrane. Interaction with ATG14 promotes translocation to autophagosomes. Expressed in dendrites and cell bodies of cerebellar Purkinje cells. - Information by UniProt
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Database links
- Entrez Gene: 8678 Human
- Omim: 604378 Human
- SwissProt: Q14457 Human
- Unigene: 716464 Human
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Alternative names
- APG6 antibody
- ATG 6 antibody
- ATG6 antibody
see all
Images
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All lanes : Anti-Beclin 1 antibody [OTI4A10] (ab118148) at 1/2000 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : BECN1 CRISPR-Cas9 edited Jurkat cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Raji cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 52 kDaFalse colour image of Western blot: Anti-Beclin 1 antibody [OTI4A10] staining at 1/2000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab118148 was shown to bind specifically to Beclin 1. A band was observed at 52 kDa in wild-type Jurkat cell lysates with no signal observed at this size in BECN1 CRISPR-Cas9 edited cell line ab283778 (BECN1 CRISPR-Cas9 edited cell lysate ab283792). The band observed in the CRISPR-Cas9 edited lysate lane above 52 kDa is likely to represent Beclin 1 with an insertion. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and BECN1 CRISPR-Cas9 edited Jurkat 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
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All lanes : 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
Lane 2 : BECN1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab118148 observed at 60 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
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 ab118148 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Beclin 1 antibody [OTI4A10] (ab118148) at 1/2000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with pCMV6-ENTRY control
Lane 2 : HEK-293T cells transfected with pCMV6-ENTRY Beclin 1
Lysates/proteins at 5 µg per lane.
Predicted band size: 52 kDa
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab118148 has been referenced in 1 publication.
- Blaszczak W et al. Fucoidan Exerts Anticancer Effects Against Head and Neck Squamous Cell Carcinoma In Vitro. Molecules 23:N/A (2018). PubMed: 30545161