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Knockout Tested Rabbit Recombinant Monoclonal ATG4B antibody. Carrier free. Suitable for WB, IHC-P and reacts with Human samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PFlow Cyt (Intra)IPICC/IFIHC-Fr
Human
Tested
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human, Rat
Dilution info
-
Notes

-

Target data

Function

Cysteine protease that plays a key role in autophagy by mediating both proteolytic activation and delipidation of ATG8 family proteins (PubMed:15169837, PubMed:15187094, PubMed:17347651, PubMed:19322194, PubMed:21177865, PubMed:22302004, PubMed:26378241, PubMed:27527864, PubMed:28633005, PubMed:28821708, PubMed:29232556, PubMed:30076329, PubMed:30443548, PubMed:30661429). Required for canonical autophagy (macroautophagy), non-canonical autophagy as well as for mitophagy (PubMed:33773106, PubMed:33909989). The protease activity is required for proteolytic activation of ATG8 family proteins: cleaves the C-terminal amino acid of ATG8 proteins MAP1LC3A, MAP1LC3B, MAP1LC3C, GABARAPL1, GABARAPL2 and GABARAP, to reveal a C-terminal glycine (PubMed:15169837, PubMed:15187094, PubMed:17347651, PubMed:19322194, PubMed:20818167, PubMed:21177865, PubMed:22302004, PubMed:27527864, PubMed:28287329, PubMed:28633005, PubMed:29458288, PubMed:30661429). 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:15169837, PubMed:15187094, PubMed:17347651, PubMed:19322194, PubMed:21177865, 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:15187094, PubMed:19322194, PubMed:28633005, PubMed:29458288, PubMed:32686895, PubMed:33909989). Catalyzes delipidation of PE-conjugated forms of ATG8 proteins during macroautophagy (PubMed:15187094, PubMed:19322194, PubMed:29458288, PubMed:32686895, PubMed:33909989). Also involved in non-canonical autophagy, a parallel pathway involving conjugation of ATG8 proteins to single membranes at endolysosomal compartments, by catalyzing delipidation of ATG8 proteins conjugated to phosphatidylserine (PS) (PubMed:33909989). Compared to other members of the family (ATG4A, ATG4C or ATG4C), constitutes the major protein for proteolytic activation of ATG8 proteins, while it displays weaker delipidation activity than other ATG4 paralogs (PubMed:29458288, PubMed:30661429). 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).

Alternative names

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal ATG4B antibody. Carrier free. Suitable for WB, IHC-P and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR28756-91
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C

Notes

ab317822 is the carrier-free version of Anti-ATG4B antibody [EPR28756-91] ab317821.

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

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.

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.

Supplementary info

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

ATG4B is a cysteine protease also known as autophagin-1 with a molecular mass of approximately 45 kDa. It is involved in the autophagy process by cleaving ATG8 family proteins. This protease facilitates the lipidation of LC3 an essential component of the autophagosome membrane. You find ATG4B expressed in various tissues especially in the brain liver and muscle where it plays important roles in cellular maintenance.

Biological function summary

ATG4B functions in the cellular autophagic machinery cleaving the C-terminal of ATG8 proteins. This processing is necessary for the conjugation to phosphatidylethanolamine and integration into autophagosomes. ATG4B is part of a larger autophagic complex that also includes other critical proteins like ATG7 and ATG3 helping prepare ATG8 proteins for autophagosome creation.

Pathways

ATG4B participates in the autophagy pathway by refining the cargo processing mechanism. It interacts with the mTOR signaling pathway which influences cell survival and growth based on nutrient availability. The ATG4B enzyme is closely related to ATG7 in the pathway as both contribute to the maturation of autophagosomes.

Associated diseases and disorders

ATG4B has significant links to cancer and neurodegenerative disorders. In cancer ATG4B activity affects tumor survival since cancer cells exploit autophagy for growth under stress conditions. In neurodegenerative diseases like Alzheimer's alterations in ATG4B activity disrupt cellular homeostasis. This disruption connects to proteins like LC3 which are pivotal in neuron survival and function.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

7 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822)

    This data was developed using Anti-ATG4B antibody [EPR28756-91] ab317821, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded (A) Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell pellet (B) ATG4B knockout HeLa cell pellet tissue labeling ATG4B with Anti-ATG4B antibody [EPR28756-91] ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on (A) Wild-type HeLa cell pellet, no staining on (B) ATG4B knockout HeLa cell pellet.

    The section was incubated with Anti-ATG4B antibody [EPR28756-91] ab317821 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822)

    This data was developed using Anti-ATG4B antibody [EPR28756-91] ab317821, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ATG4B with Anti-ATG4B antibody [EPR28756-91] ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Low expression: very weak staining on human colon (PMID: 24991826).

    The section was incubated with Anti-ATG4B antibody [EPR28756-91] ab317821 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822)

    This data was developed using Anti-ATG4B antibody [EPR28756-91] ab317821, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's lymphoma tissue labeling ATG4B with Anti-ATG4B antibody [EPR28756-91] ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on human non-Hodgkin's lymhoma.

    The section was incubated with Anti-ATG4B antibody [EPR28756-91] ab317821 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822)

    This data was developed using Anti-ATG4B antibody [EPR28756-91] ab317821, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human colorectal carcinoma tissue labeling ATG4B with Anti-ATG4B antibody [EPR28756-91] ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on human colorectal carcinoma (PMID: 24991826).

    The section was incubated with Anti-ATG4B antibody [EPR28756-91] ab317821 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822)

    This data was developed using Anti-ATG4B antibody [EPR28756-91] ab317821, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling ATG4B with Anti-ATG4B antibody [EPR28756-91] ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on human tonsil.

    The section was incubated with Anti-ATG4B antibody [EPR28756-91] ab317821 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a 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.

  • Western blot - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822), expandable thumbnail

    Western blot - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822)

    This data was developed using Anti-ATG4B antibody [EPR28756-91] ab317821, the same antibody clone in a different buffer formulation.

    In Western blot, Anti-ATG4B antibody [EPR28756-91] ab317821 was shown to bind specifically to ATG4B. A band was observed at 44 kDa in wild-type Hela cell lysates whereas loss of signal was observed in the ATG4B knockout cell line (Human ATG4B knockout HeLa cell line ab265814/knockout lysate Human ATG4B knockout HeLa cell lysate ab257364).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    Exposure time: Lanes 1-2: 15 seconds, Lanes 3-5: 48 seconds

    All lanes: Western blot - Anti-ATG4B antibody [EPR28756-91] (Anti-ATG4B antibody [EPR28756-91] ab317821) at 1/1000 dilution

    Lane 1: Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 2: ATG4B knockout HeLa whole cell lysate at 20 µg with NFDM/TBST

    Lane 3: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 4: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg with NFDM/TBST

    Lane 5: Ramos (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg with NFDM/TBST

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 44 kDa, 36 kDa

  • Western blot - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822), expandable thumbnail

    Western blot - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (ab317822)

    This data was developed using Anti-ATG4B antibody [EPR28756-91] ab317821, the same antibody clone in a different buffer formulation.

    All lanes: Western blot - Anti-ATG4B antibody [EPR28756-91] (Anti-ATG4B antibody [EPR28756-91] ab317821) at 1/1000 dilution

    All lanes: Human tonsil tissue lysate at 20 µg with NFDM/TBST

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Developed using the ECL technique.

    Observed band size: 44 kDa

    Exposure time: 81s

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Product protocols

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