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Knockout Tested Rabbit Recombinant Monoclonal ATG4B antibody. 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] (AB317821), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (AB317821), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (AB317821), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (AB317821), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (AB317821), expandable thumbnail

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.

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

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

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

-

Associated Products

Select an associated product type

1 products for Alternative Version

    Target data

    Function

    Cysteine protease required for the cytoplasm to vacuole transport (Cvt) and autophagy. Cleaves the C-terminal amino acid of ATG8 family proteins MAP1LC3, GABARAPL1, GABARAPL2 and GABARAP, to reveal a C-terminal glycine. 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. Has also an activity of delipidating enzyme for the PE-conjugated forms.

    Alternative names

    Cysteine protease ATG4B, AUT-like 1 cysteine endopeptidase, Autophagin-1, Autophagy-related cysteine endopeptidase 1, Autophagy-related protein 4 homolog B, hAPG4B, APG4B, KIAA0943, AUTL1, ATG4B

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

    Alternative names

    Cysteine protease ATG4B, AUT-like 1 cysteine endopeptidase, Autophagin-1, Autophagy-related cysteine endopeptidase 1, Autophagy-related protein 4 homolog B, hAPG4B, APG4B, KIAA0943, AUTL1, ATG4B

    Key facts

    Isotype

    IgG

    Form

    Liquid

    Clonality

    Monoclonal

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

    EPR28756-91

    Purification technique

    Affinity purification Protein A

    Concentration
    Loading...

    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

    Avoid freeze / thaw cycle

    Notes

    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.

    Supplementary info

    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] (ab317821), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (ab317821)

      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 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 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] (ab317821), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (ab317821)

      Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ATG4B with 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 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] (ab317821), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (ab317821)

      Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's lymphoma tissue labeling ATG4B with 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 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] (ab317821), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (ab317821)

      Immunohistochemical analysis of paraffin-embedded Human colorectal carcinoma tissue labeling ATG4B with 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 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] (ab317821), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] (ab317821)

      Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling ATG4B with 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 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] (ab317821), expandable thumbnail

      Western blot - Anti-ATG4B antibody [EPR28756-91] (ab317821)

      In Western blot, 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] (ab317821) at 1/1000 dilution

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

      Lane 2: ATG4B knockout HeLa whole cell lysate at 20 µg

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

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

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

      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] (ab317821), expandable thumbnail

      Western blot - Anti-ATG4B antibody [EPR28756-91] (ab317821)

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

      All lanes: Human tonsil tissue lysate at 20 µg

      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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