Autophagy Antibody Panel Sampler Panel (ab228525) contains multiple trial-sized versions of clones against APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, and LC3B specifically selected for high performance in various applications.
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- Naunyn-Schmiedeberg's archives of pharmacology 396:1105-11152023Doxorubicin-induced acute cardiotoxicity is associated with increased oxidative stress, autophagy, and inflammation in a murine model.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPatricia Lorena Dulf,Mihaela Mocan,Camelia Alexandra Coadă,Daniel Vasile Dulf,Remus Moldovan,Ioana Baldea,Anca-Daniela Farcas,Dan Blendea,Adriana Gabriela FilipPubMed 36645429
- Aging 12:21687-217052020Dexmedetomidine inhibits inflammatory response and autophagy through the circLrp1b/miR-27a-3p/Dram2 pathway in a rat model of traumatic brain injury.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHengchang Li et. al.PubMed 33147167
Key facts
- Reactive species
- Mouse, Rat, Human
- Target
- ATG5
(See target data below)
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Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application WB | Reactivity Reacts | Dilution info - | Notes - |
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Target data
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. 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. As part of the ATG8 conjugation system with ATG12 and ATG16L1, required for recruitment of LRRK2 to stressed lysosomes and induction of LRRK2 kinase activity in response to lysosomal stress (By similarity). 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. (Microbial infection) May act as a proviral factor. In association with ATG12, negatively regulates the innate antiviral immune response by impairing the type I IFN production pathway upon vesicular stomatitis virus (VSV) infection (PubMed:17709747). Required for the translation of incoming hepatitis C virus (HCV) RNA and, thereby, for initiation of HCV replication, but not required once infection is established (PubMed:19666601).
Additional Targets
ATG16L1, ATG4B, ATG9A, BECN1, MAP1LC3B
Alternative names
APG5L, ASP, ATG5, Autophagy protein 5, APG5-like, Apoptosis-specific protein
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Autophagy Antibody Panel Sampler Panel (ab228525) contains multiple trial-sized versions of clones against APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, and LC3B specifically selected for high performance in various applications.
Key facts
- Reactive species
- Mouse, Rat, Human
- Target
- ATG5
- Applications
- WB
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- Multi
- Appropriate long-term storage conditions
- Multi
- Storage information
- Please refer to protocols
Notes
Autophagy Antibody Panel Sampler Panel (ab228525) contains multiple trial-sized versions of clones against APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, and LC3B specifically selected for high performance in various applications. This panel contains 6 recombinant rabbit monoclonal antibodies, 5 of which are knock-out validated. They are provided as a sampler panel to allow you to easily evaluate each in your required applications.
For guidelines on how to use each antibody within the panel, please consult the individual datasheet for each antibody.
Panel contains:
- Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5 (20 μL) ab108327
- Rabbit monoclonal [EPR15638] to ATG16L1 - N-terminal (20 μL) ab187671
- Rabbit monoclonal [EPR6436(2)] to ATG4B (20 μL) ab154843
- Rabbit monoclonal [EPR2450(2)] to ATG9A (20 μL) ab108338
- Rabbit monoclonal [EPR19662] to Beclin 1 (20 μL) ab207612
- Rabbit monoclonal [EPR18709] to LC3B (20 μL) ab192890
Explore our range of antibody sample panels designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format.
Carrier-free formulations of our recombinant antibodies are also available for easy conjugation to labels of your choice and for multiplex applications. Use our intuitive search and select carrier-free or your label of choice. For bespoke conjugations or large volumes email bespoke@abcam.com.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Beclin 1 also known as ATG6 or VPS30 is a central player in the autophagy process. This protein has a molecular mass of approximately 60 kDa and is widely expressed in a variety of tissues. Beclin 1 forms a core component of the class III phosphatidylinositol 3-kinase (PI3K) complex and helps to initiate the formation of pre-autophagosomal structures. Associated proteins like APG5L/ATG5 ATG16L1 ATG9A LC3B and ATG4B also play important roles in the regulation of autophagy. Particularly LC3B serves as an important autophagy marker often used to monitor autophagosomes through autophagy antibodies in experimental settings.
- Biological function summary
These proteins coordinate to regulate and execute autophagy a process that cleans up damaged cellular components and helps maintain cellular homeostasis. They assemble into a complex network that orchestrates the formation and maturation of autophagosomes effectively marking them for lysosomal degradation. Beclin 1 interacts with multiple proteins like Atg14 and UVRAG to form different PI3K complexes leading to nucleation of the autophagic membrane. ATG5 ATG12 and ATG16L1 work in a conjugation system that expands the autophagosome membrane while the proteolytic processing of LC3B is essential for autophagosome closure.
- Pathways
The autophagy process plays a significant role in cellular pathways like nutrient sensing and stress response. The AMPK/mTOR pathway for example tightly regulates autophagy in response to cellular energy status. AMPK activates autophagy by inhibiting mTOR signaling allowing initiation of autophagosome formation. Proteins such as ATG9A are integral for the trafficking of autophagy-related vesicles affecting the recruitment and movement of membrane source for developing autophagosomes. The balance between PI3K-AKT signaling and mTOR is important for the coordination between cell growth and the autophagic activity mediated by these proteins.
- Associated diseases and disorders
Dysregulated autophagy pathways involving Beclin 1 and associated components like LC3B and ATG5 are linked to cancer and neurodegenerative diseases such as Alzheimer's. Alterations in autophagy can disrupt cellular homeostasis facilitating the development of malignancies through the evasion of cell death. In neurodegenerative contexts impaired autophagy leads to the accumulation of toxic proteins exacerbating disease progression. The interplay between Beclin 1 and proteins like Bcl-2 also links it to apoptosis suggesting that a delicate balance between autophagy and apoptosis is important in preventing these disorders.
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5 product images
Western blot - Autophagy Marker (APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, LC3B) Antibody Sampler Panel (ab228525)
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 - Anti-APG5L/ATG5 antibody [EPR1755(2)] ab108327 observed at 52 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-APG5L/ATG5 antibody [EPR1755(2)] ab108327 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. Anti-APG5L/ATG5 antibody [EPR1755(2)] ab108327 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (Anti-APG5L/ATG5 antibody [EPR1755(2)] ab108327)
Predicted band size: 32 kDa
Immunocytochemistry/ Immunofluorescence - Autophagy Marker (APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, LC3B) Antibody Sampler Panel (ab228525)
LC3B immunofluorescence staining of HeLa cells using rabbit anti-LC3B antibody
Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab192890 staining LC3B in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells +/- Chloroquine (50μM 24 hours).The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab192890 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594) at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Alexa Fluor® 488 (Alexa Fluor® 647 Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab225383) and Alexa Fluor® 647 (Alexa Fluor® 488 Anti-LC3B antibody [EPR18709] - Autophagosome Marker ab225382) conjugated versions are available for this clone.
Western blot - Autophagy Marker (APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, LC3B) Antibody Sampler Panel (ab228525)
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: ATG4B knockout HAP1 cell lysate (20 μg)Lanes 1 - 2: Merged signal (red and green). Green - Anti-ATG4B antibody [EPR6436(2)] ab154843 observed at 47 kDa. Red - loading control, ab18058, observed at 130 kDa.
Anti-ATG4B antibody [EPR6436(2)] ab154843 was shown to specifically react with ATG4B when ATG4B knockout samples were used. Wild-type and ATG4B knockout samples were subjected to SDS-PAGE. Anti-ATG4B antibody [EPR6436(2)] ab154843 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-ATG4B antibody [EPR6436(2)] (Anti-ATG4B antibody [EPR6436(2)] ab154843)
Predicted band size: 44 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Autophagy Marker (APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, LC3B) Antibody Sampler Panel (ab228525)
Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling ATG16L1 with Anti-ATG16L1 antibody [EPR15638] - N-terminal ab187671 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Autophagy Marker (APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, LC3B) Antibody Sampler Panel (ab228525)
Anti-ATG9A antibody [EPR2450(2)] ab108338, at 1/100, staining ATG9A in paraffin-embedded Human colon tissue by Immunohistochemistry.
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