Recombinant Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)
![Western blot - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)](https://www.abcam.com/ps/products/227/ab227084/Images/ab227084-512520-anti-apg5latg5-antibody-epr4797-bsa-and-azide-free-western-blot-wildtype-thp1-atg5-knockout-thp1-u87-mg-hela.jpg "Western blot - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4797] to APG5L/ATG5 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free
See all APG5L/ATG5 primary antibodies -
Description
Rabbit monoclonal [EPR4797] to APG5L/ATG5 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- THP1, Raji, HeLa, HT1080 and Human fetal kidney lysates; Human breast carcinoma and Human kidney tissues.
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General notes
ab227084 is the carrier-free version of ab109490.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4797 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
- Alexa Fluor® 488 Anti-APG5L/ATG5 antibody [EPR4797] (ab206714)
- Alexa Fluor® 647 Anti-APG5L/ATG5 antibody [EPR4797] (ab206715)
- Alexa Fluor® 555 Anti-APG5L/ATG5 antibody [EPR4797] (ab211977)
- PE Anti-APG5L/ATG5 antibody [EPR4797] (ab306274)
- HRP Anti-APG5L/ATG5 antibody [EPR4797] (ab306276)
- Alexa Fluor® 594 Anti-APG5L/ATG5 antibody [EPR4797] (ab310480)
- Alexa Fluor® 568 Anti-APG5L/ATG5 antibody [EPR4797] (ab312486)
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Conjugation kits
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Isotype control
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KO cell lines
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227084 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 |
|---|---|---|
| Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
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| WB |
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.
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| IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Notes |
|---|
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa. |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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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Western blot - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)All lanes : Anti-APG5L/ATG5 antibody [EPR4797] (ab109490) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : ATG5 knockout THP-1 cell lysate
Lane 3 : U-87 MG cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-APG5L/ATG5 antibody [EPR4797] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109490 was shown to bind specifically to APG5L/ATG5. A band was observed at 50 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG5 knockout cell line ab277835 (knockout cell lysate ab290722). To generate this image, wild-type and ATG5 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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.
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![Western blot - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)](https://www.abcam.com/ps/products/227/ab227084/Images/ab227084-288826-ab109490KOWB.jpg)
Western blot - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)This WB data was generated using the same anti-APG5/ATG5 antibody clone, EPR4797, in a different buffer formulation (cat# ab109490).
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: APG5L/ATG5 knockout HAP1 cell lysate (20 µg)
Lane 3: Raji cell lysate (20 µg)
Lane 4: Jeg-3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109490 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109490 was shown to specifically react with APG5L/ATG5 when APG5L/ATG5 knockout samples were used. Wild-type and APG5L/ATG5 knockout samples were subjected to SDS-PAGE. ab109490 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging. -
![Flow Cytometry (Intracellular) - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)](https://www.abcam.com/ps/products/227/ab227084/Images/ab227084-4-ab1094906ab109490FC1.jpg)
Flow Cytometry (Intracellular) - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling APG5L/ATG5 with purified ab109490 at 1/250 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109490).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)](https://www.abcam.com/ps/products/227/ab227084/Images/ab227084-3-APG5LATG5Primaryantibodiesab1094904.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)ab109490, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human breast carcinoma by Immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109490).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)](https://www.abcam.com/ps/products/227/ab227084/Images/ab227084-2-APG5LATG5Primaryantibodiesab1094905.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)ab109490, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human kidney by Immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109490).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)](https://www.abcam.com/ps/products/227/ab227084/Images/ab227084-5-benefits-of-recombinant-antibodies.png)
Anti-APG5L/ATG5 antibody [EPR4797] - BSA and Azide free (ab227084)
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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Datasheet download
Certificate of Compliance
References (4)
ab227084 has been referenced in 4 publications.
- Chen W et al. Hyperbaric oxygen protects against myocardial ischemia-reperfusion injury through inhibiting mitochondria dysfunction and autophagy. Mol Med Rep 22:4254-4264 (2020). PubMed: 32901878
- He G et al. Gadd45b prevents autophagy and apoptosis against rat cerebral neuron oxygen-glucose deprivation/reperfusion injury. Apoptosis 21:390-403 (2016). WB ; Rat . PubMed: 26882903
- Zhang L et al. Reprogramming towards anabolism impedes degeneration in a preclinical model of retinitis pigmentosa. Hum Mol Genet : (2016). PubMed: 27516389
- Pla A et al. TLR4 mediates the impairment of ubiquitin-proteasome and autophagy-lysosome pathways induced by ethanol treatment in brain. Cell Death Dis 5:e1066 (2014). WB ; Mouse . PubMed: 24556681
