Recombinant Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [EPR1755(2)] to APG5L/ATG5 - Chimeric
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric)
See all APG5L/ATG5 primary antibodies -
Description
Mouse monoclonal [EPR1755(2)] to APG5L/ATG5 - Chimeric -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HAP1 whole cell lysate. Raji, C6, RAW264.7, PC-12 and NIH/3T3 whole cell lysate. Mouse brain, heart, kidney and spleen tissue lysate. Rat brain and heart tissue lysate. IHC-P: Human ovarian cancer tissue. Parental HAP1 cell pellet. Mouse and rat colon tissue.
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General notes
ab307842 is a mouse monoclonal chimeric antibody.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1755(2) -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab307842 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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IHC-P |
1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 32 kDa.
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Notes |
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IHC-P
1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 32 kDa. |
Target
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Function
Involved in autophagic vesicle formation. Conjugation with ATG12, through a ubiquitin-like conjugating system involving ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme, is essential for its function. The ATG12-ATG5 conjugate acts as an E3-like enzyme which is required for lipidation of ATG8 family proteins and their association to the vesicle membranes. Involved in mitochondrial quality control after oxidative damage, and in subsequent cellular longevity. The ATG12-ATG5 conjugate also negatively regulates the innate antiviral immune response by blocking the type I IFN production pathway through direct association with RARRES3 and MAVS. Also plays a role in translation or delivery of incoming viral RNA to the translation apparatus. Plays a critical role in multiple aspects of lymphocyte development and is essential for both B and T lymphocyte survival and proliferation. Required for optimal processing and presentation of antigens for MHC II. Involved in the maintenance of axon morphology and membrane structures, as well as in normal adipocyte differentiation. Promotes primary ciliogenesis through removal of OFD1 from centriolar satellites and degradation of IFT20 via the autophagic pathway.
May play an important role in the apoptotic process, possibly within the modified cytoskeleton. Its expression is a relatively late event in the apoptotic process, occurring downstream of caspase activity. Plays a crucial role in IFN-gamma-induced autophagic cell death by interacting with FADD. -
Tissue specificity
Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells. -
Sequence similarities
Belongs to the ATG5 family. -
Post-translational
modificationsConjugated to ATG12; which is essential for autophagy, but is not required for association with isolation membrane.
Acetylated by EP300. -
Cellular localization
Cytoplasm. Preautophagosomal structure membrane. Colocalizes with nonmuscle actin. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed (By similarity). Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme. - Information by UniProt
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Database links
- Entrez Gene: 9474 Human
- Entrez Gene: 11793 Mouse
- Entrez Gene: 365601 Rat
- Omim: 604261 Human
- SwissProt: Q9H1Y0 Human
- SwissProt: Q99J83 Mouse
- SwissProt: Q3MQ06 Rat
- Unigene: 486063 Human
see all -
Alternative names
- APG 5 antibody
- APG 5L antibody
- APG5 antibody
see all
Images
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All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : APG5L/ATG5 knockout HAP1 whole cell lysate
Lane 3 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
False colour image of Western blot: Anti-APG5L/ATG5 antibody [EEPR1755(2)] (ab307842) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (ab181602) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab307842 was shown to bind specifically to APG5L/ATG5 . A band was observed at 55 kDa in wild-type HAP1 cell lysates with no signal observed at this size in APG5L/ATG5 knockout cell line. To generate this image, wild-type and APG5L/ATG5 knockout HAP1 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 (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution. -
All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842) at 1/1000 dilution
Lane 1 : HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Mouse brain tissue lysate
Lane 4 : Mouse heart tissue lysate
Lane 5 : Mouse kidney tissue lysate
Lane 6 : Mouse spleen tissue lysate
Lane 7 : Rat brain tissue lysate
Lane 8 : Rat heart tissue lysate
Lane 9 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 10 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 11 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG Fc (HRP) preadsorbed (ab98717) at 1/20000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?Exposure time: Lanes 1-3: 48 seconds;Lanes 4-9: 92 seconds;Lanes 10-13: 37 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842)
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling APG5L/ATG5 with ab307842 at 1/500 (1.66 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Cytoplasmic staining on rat colon.
The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842)
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling APG5L/ATG5 with ab307842 at 1/500 (1.66 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Cytoplasmic staining on mouse colon.
The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842)
Immunohistochemical analysis of paraffin-embedded cell pellets labeling APG5L/ATG5 with ab307842 at 1/2000 dilution (0.415 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Cytoplasmic staining on (A) parental HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet, no staining on (B) APG5L/ATG5 knockout HAP1 (HO010083) cell pellet.
The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] - Mouse IgG1 (Chimeric) (ab307842)
Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling APG5L/ATG5 with ab307842 at 1/500 (1.66 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Cytoplasmic staining on human ovarian cancer.
The section was incubated with ab307842 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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 (0)
ab307842 has not yet been referenced specifically in any publications.