Validated using a knockout cell line
Recombinant
RabMAb

Anti-ATG4B antibody [EPR16572] (ab199537)

Overview

  • Product name
    Anti-ATG4B antibody [EPR16572]
    See all ATG4B primary antibodies
  • Description
    Rabbit monoclonal [EPR16572] to ATG4B
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IPmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Recombinant fragment within Human ATG4B aa 1-150. The exact sequence is proprietary.
    Database link: Q9Y4P1

  • Positive control
    • WB: 293T, Ramos, HeLa, Jurkat and PC-12 whole cell lysates; Rat brain lysate. IP: Jurkat and Ramos whole cell lysates.
  • General 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 rabbit monoclonal antibody.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab199537 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
IP 1/50.

Target

  • Function
    Cysteine protease required for autophagy, which cleaves the C-terminal part of either MAP1LC3, GABARAPL2 or GABARAP, allowing the liberation of form I. A subpopulation of form I is subsequently converted to a smaller form (form II). Form II, with a revealed C-terminal glycine, is considered to be the phosphatidylethanolamine (PE)-conjugated form, and has the capacity for the binding to autophagosomes.
  • Tissue specificity
    Mainly expressed in the skeletal muscle, followed by brain, heart, liver and pancreas.
  • Sequence similarities
    Belongs to the peptidase C54 family.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • APG4 autophagy 4 homolog B antibody
    • APG4B antibody
    • ATG4 autophagy related 4 homolog B (S. cerevisiae) antibody
    • ATG4B antibody
    • ATG4B_HUMAN antibody
    • AUT like 1 cysteine endopeptidase antibody
    • AUT-like 1 cysteine endopeptidase antibody
    • AUTL1 antibody
    • Autophagin 1 antibody
    • Autophagin-1 antibody
    • Autophagy related 4B cysteine peptidase antibody
    • Autophagy related cysteine endopeptidase 1 antibody
    • Autophagy related protein 4 homolog B antibody
    • Autophagy-related cysteine endopeptidase 1 antibody
    • Autophagy-related protein 4 homolog B antibody
    • Cysteine protease ATG4B antibody
    • hAPG4B antibody
    • MGC1353 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ATG4B knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Jurkat cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab199537 observed at 44 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab199537 was shown to specifically react with ATG4B when ATG4B knockout samples were used. Wild-type and ATG4B knockout samples were subjected to SDS-PAGE. ab199537 and ab8245 (loading control to GAPDH) were diluted 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 ab216773 and 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 : Anti-ATG4B antibody [EPR16572] (ab199537) at 1/1000 dilution

    Lane 1 : 293T (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 2 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 44 kDa
    Observed band size: 44 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-ATG4B antibody [EPR16572] (ab199537) at 1/1000 dilution

    Lane 1 : Rat brain lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 44 kDa
    Observed band size: 44 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • ATG4B was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab199537 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab199537 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Jurkat whole cell lysate 10ug (Input). Lane 2: ab199537 IP in Jurkat whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199537 in Jurkat whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • ATG4B was immunoprecipitated from 1mg of Ramos (Human Burkitt's lymphoma cell line) whole cell lysate with ab199537 at 1/60 dilution. Western blot was performed from the immunoprecipitate using ab199537 at 1/500 dilution (Panel A) or ab154843 at 1/500 dilution (Panel B). Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Ramos whole cell lysate 10ug (Input). Lane 2: ab199537 IP in Ramos whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199537 in Ramos whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: Panel A and B 30 seconds.

References

ab199537 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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