Name: Anti-APG5L/ATG5 antibody [EPR1755(2)]
SKU: ab108327
Rating: 4 (7 reviews)

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) is a rabbit monoclonal antibody that is used to detect APG5L/ATG5 in Western Blot, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with APG5L/ATG5 knockout cell line validation

Images

Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (AB108327), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	IHC-P
Human	Expected	Tested	Tested	Tested
Mouse	Expected	Tested	Expected	Expected
Rat	Tested	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/10 - 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/10000	Notes -
Species Rat	Dilution info 1/1000 - 1/10000	Notes -
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes Antigen retrieval is recommended. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

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Target data

See full target information ATG5

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).

Alternative names

APG5L, ASP, ATG5, Autophagy protein 5, APG5-like, Apoptosis-specific protein

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR1755(2)
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

What is this antibody validated in?

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), in Human, Mouse, Rat samples.

What is the molecular weight of APG5L/ATG5?

Anti-APG5L/ATG5 [EPR1755(2)] (ab108327) specifically detects a band for APG5L/ATG5 (UniProt: Q9H1Y0) at a molecular weight of 32kDa.

Recommended positive controls

Raji, HeLa, HT-1080, Human fetal kidney, C6, Raw264.7, PC-12 and NIH3T3 cell lysates; Human hepatocellular carcinoma and Human ovarian adenocarcinoma tissue

Trusted by the scientific community

Anti-APG5L/ATG5 [EPR1755(2)] (ab108327) was first used in a scientific publication in 2012 and has been cited over 190 times in peer-reviewed journals.

Reviewed by scientists

Anti-APG5L/ATG5 [EPR1755(2)] (ab108327) has over 5 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies. Specificity confirmed

The specificity of Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) has been confirmed by Western blot testing in ATG5 Knockout HAP1 cell line, ab277835.

Other related products

We have a range of other formats of antibody clone [EPR1755(2)] also available for your convenience:
ab108327, HRP - ab199560, Carrier free - ab221604, ab307842, Carrier free - ab307843

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

APG5L commonly known as ATG5 or ATG5 protein plays a significant role in autophagy. This molecular component has a molecular weight of approximately 32 kDa. ATG5 is expressed in multiple tissue types with a higher expression profile in immune-related cells. The protein is essential for the formation of the autophagosome as it conjugates with ATG12 to form a complex that is critical for autophagosome membrane elongation.

Biological function summary

ATG5 plays an important part in the cellular process of autophagy affecting the degradation and recycling of cellular components. It is a part of a complex with ATG12 and ATG16L1 which facilitates the membrane building of the autophagosome. This protein is important for maintaining cellular homeostasis allowing the removal of damaged proteins and organelles and is involved in responses to stress and starvation.

Pathways

ATG5 is an important component in the autophagy pathway and is involved in cellular maintenance processes. This pathway is vital for the regulation of cell growth and metabolism. ATG5 works closely with other autophagy-related proteins such as LC3. The ATG5-ATG12 complex helps in the lipidation of LC3 which is critical for autophagosome maturation connecting its function to the mTOR signaling pathway that regulates many cellular processes.

Associated diseases and disorders

Disruption in ATG5 function is linked to neurodegenerative diseases and cancer. For instance dysregulation of autophagy due to ATG5 dysfunction may result in the progression of neurodegenerative disorders like Alzheimer's disease. Additionally in certain cancers altered autophagy can influence the survival of cancer cells. The interplay between ATG5 and other proteins like p53 and Bcl-2 is under investigation to understand its role in these diseases.

Product promise

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12 product images

Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
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 - ab108327 observed at 52 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
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. 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)] (ab108327)
Predicted band size: 32 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human ovarian adenocarcinoma tissue by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Immunofluorescence staining of MCF7 cells with purified ab108327 at a working dilution of 1/150, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab108327 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution
Lane 1: C6 cell lysate at 20 µg
Lane 2: PC-12 cell lysate at 20 µg
Lane 3: NIH/3T3 cell lysate at 20 µg
Secondary
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution
All lanes: Raji cell lysate at 20 µg
Secondary
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/10000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa
Immunoprecipitation - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Recombinant Human MED3 protein (denatured) ab180327 (purified) at 1/20 immunoprecipitating CRSP8 in 10 μg PC-12 whole cell lysate (Lanes 1 and 2, observed at 55 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1500 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Predicted band size: 32 kDa
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution
All lanes: HT-1080 cell lysate at 20 µg
Secondary
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution
All lanes: human fetal kidney at 10 µg
Secondary
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 102 kDa, 32 kDa
Observed band size: 32 kDa, 55 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified Recombinant Human MED3 protein (denatured) ab180327 at a working dilution of 1/150. The secondary antibody used is HRP goat anti-rabbit IgG H&L (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Secondary antibody - anti-rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab6721)
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/1000 dilution
Lane 1: Raji cell lysate at 10 µg
Lane 2: HeLa cell lysate at 10 µg
Lane 3: HT-1080 cell lysate at 10 µg
Lane 4: Human fetal kidney cell lysate at 10 µg
Lane 5: C6 cell lysate at 10 µg
Lane 6: Raw264.7 cell lysate at 10 µg
Lane 7: PC-12 cell lysate at 10 µg
Lane 8: NIH3T3 cell lysate at 10 µg
Predicted band size: 32 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human hepatocellular carcinoma tissue by Immunohistochemistry.
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)
False colour image of Western blot: Anti-APG5L/ATG5 antibody [EPR1755(2)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab108327 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 Human ATG5 knockout THP-1 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.
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: ATG5 knockout THP-1 cell lysate at 20 µg
Lane 3: U-87 MG cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Secondary
Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 50 kDa