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AB247630

Anti-ATG4A antibody [EPR4122] - BSA and Azide free

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Rabbit Recombinant Monoclonal ATG4A antibody. Carrier free. Suitable for IHC-P, WB and reacts with Human samples.

View Alternative Names

APG4A, AUTL2, ATG4A, Cysteine protease ATG4A, AUT-like 2 cysteine endopeptidase, Autophagy-related cysteine endopeptidase 2, Autophagy-related protein 4 homolog A, Autophagin-2, HsAPG4A, hAPG4A

4 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)

This data was developed using ab108322, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue using ab108322 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)

This data was developed using ab108322, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human brain tissue using ab108322 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Western blot - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)
  • WB

Unknown

Western blot - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)

This data was developed using ab108322, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ATG4A antibody [EPR4122] (<a href='/en-us/products/primary-antibodies/atg4a-antibody-epr4122-ab108322'>ab108322</a>) at 1/1000 dilution

Lane 1:

Fetal liver tissue lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

Fetal brain tissue lysate at 10 µg

Secondary

All lanes:

Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 45 kDa

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Western blot - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)
  • WB

Lab

Western blot - Anti-ATG4A antibody [EPR4122] - BSA and Azide free (AB247630)

This data was developed using the same antibody clone in a different buffer formulation (ab108322).

Lanes 1 - 4 : Merged signal (red and green). Green - ab108322 observed at 45 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab108322 was shown to react with ATG4A in wild-type HeLa cells in Western blot with loss of signal observed in ATG4A knockout cell line ab265738 (ATG4A knockout cell lysate ab257846). Wild-type HeLa and ATG4A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab108322 and ab8245 (Mouse anti-GAPDH antibody [6C5]) 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-ATG4A antibody [EPR4122] (<a href='/en-us/products/primary-antibodies/atg4a-antibody-epr4122-ab108322'>ab108322</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ATG4A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ATG4A knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atg4a-knockout-hela-cell-line-ab265738'>ab265738</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 45 kDa

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Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR4122

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, IHC-P

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p>Perform antigen retrieval.</p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Mouse": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Rat": { "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" } } }
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Product details

ab247630 is the carrier-free version of ab108322.

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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

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

ATG4A also known as Autophagin 1 is a cysteine protease with a mass of approximately 52 kDa. It plays a critical role in the autophagy pathway by cleaving LC3 and other Atg8 homologs facilitating their conjugation to phosphatidylethanolamine on the autophagosome membrane. This protein is expressed in various tissues including the liver and skeletal muscle indicating its involvement in diverse physiological processes.
Biological function summary

ATG4A contributes essential functions to cellular homeostasis by participating in autophagosome formation. As part of the group of Atg proteins it collaborates with other members to modulate the autophagy machinery. ATG4A's enzymatic activity provides the necessary steps for processing LC3 transforming it into a form that allows lipidation and incorporation into autophagic vesicles.

Pathways

ATG4A is integral to the autophagy pathway a catabolic process important for cell survival under stress conditions. It interacts with proteins such as ATG7 and ATG3 within this pathway playing a significant role in the early stages of autophagosome maturation. The interactions ensure proper recycling of cellular materials and its efficient function impact cellular metabolism and stress responses.

Disruptions in ATG4A function have been linked to neurodegenerative diseases and cancer. Aberrant autophagy processes involving ATG4A can lead to protein aggregation disorders such as Alzheimer’s disease where proper proteostasis is compromised. Also in certain cancers altered ATG4A activity can influence tumor growth often through modulation of metabolic pathways and interactions with proteins like Beclin 1 which regulate autophagic responses.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Cysteine protease that plays a key role in autophagy by mediating both proteolytic activation and delipidation of ATG8 family proteins (PubMed : 12473658, PubMed : 15169837, PubMed : 17347651, PubMed : 21177865, PubMed : 21245471, PubMed : 22302004, PubMed : 32732290). The protease activity is required for proteolytic activation of ATG8 family proteins : cleaves the C-terminal amino acid of ATG8 proteins to reveal a C-terminal glycine (PubMed : 12473658, PubMed : 15169837, PubMed : 17347651, PubMed : 21177865, PubMed : 21245471, PubMed : 22302004). Exposure of the glycine at the C-terminus is essential for ATG8 proteins conjugation to phosphatidylethanolamine (PE) and insertion to membranes, which is necessary for autophagy (PubMed : 12473658, PubMed : 15169837, PubMed : 17347651, PubMed : 21177865, PubMed : 21245471, PubMed : 22302004). Preferred substrate is GABARAPL2 followed by MAP1LC3A and GABARAP (PubMed : 12473658, PubMed : 15169837, PubMed : 17347651, PubMed : 21177865, PubMed : 21245471, PubMed : 22302004). Protease activity is also required to counteract formation of high-molecular weight conjugates of ATG8 proteins (ATG8ylation) : acts as a deubiquitinating-like enzyme that removes ATG8 conjugated to other proteins, such as ATG3 (PubMed : 31315929, PubMed : 33773106). In addition to the protease activity, also mediates delipidation of ATG8 family proteins (PubMed : 29458288, PubMed : 33909989). Catalyzes delipidation of PE-conjugated forms of ATG8 proteins during macroautophagy (PubMed : 29458288, PubMed : 33909989). Compared to ATG4B, the major protein for proteolytic activation of ATG8 proteins, shows weaker ability to cleave the C-terminal amino acid of ATG8 proteins, while it displays stronger delipidation activity (PubMed : 29458288). Involved in phagophore growth during mitophagy independently of its protease activity and of ATG8 proteins : acts by regulating ATG9A trafficking to mitochondria and promoting phagophore-endoplasmic reticulum contacts during the lipid transfer phase of mitophagy (PubMed : 33773106).
See full target information ATG4A
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Product promise

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