Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ATG4B antibody [EPR6436(2)] (ab154843)

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Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
  • Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
  • Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
  • Flow Cytometry (Intracellular) - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
  • Anti-ATG4B antibody [EPR6436(2)] (ab154843)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6436(2)] to ATG4B
  • Suitable for: Flow Cyt (Intra), WB
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-ATG4B antibody [EPR6436(2)]
    See all ATG4B primary antibodies

  • Description

    Rabbit monoclonal [EPR6436(2)] to ATG4B

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WBmore details
    Unsuitable for: ICC/IF,IHC-P or IP

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat

  • Immunogen

    Synthetic peptide within Human ATG4B. The exact sequence is proprietary.

  • Positive control

    • Recombinant Human ATG4B protein (ab123195) can be used as a positive control in WB. Ramos, 293T, Jurkat and HeLa cell lysates; Jurkat cells.

  • General notes

    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 production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab154843 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
Flow Cyt (Intra)
1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 44 kDa.
Notes
Flow Cyt (Intra)
1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  
WB
1/1000 - 1/10000. Predicted molecular weight: 44 kDa.
Application notes
Is unsuitable for ICC/IF,IHC-P or IP.

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

    see all

  • 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

  • Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
    Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
    All lanes : Anti-ATG4B antibody [EPR6436(2)] (ab154843) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG4B knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 44 kDa
    Observed band size: 47 kDa why is the actual band size different from the predicted?



    Lanes 1 - 2: Merged signal (red and green). Green - ab154843 observed at 47 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab154843 was shown to react with ATG4B in wild-type HEK293T cells in Western blot with loss of signal observed in ATG4B knockout cell line ab265814 (ATG4B knockout cell lysate ab257364). Wild-type and ATG4B knockout HEK293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab154843 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.

  • Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
    Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ATG4B knockout HAP1 cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab154843 observed at 47 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab154843 was shown to specifically react with ATG4B when ATG4B knockout samples were used. Wild-type and ATG4B knockout samples were subjected to SDS-PAGE. ab154843 and ab18058 (loading control to Vinculin) were diluted at 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.

  • Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
    Western blot - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
    All lanes : Anti-ATG4B antibody [EPR6436(2)] (ab154843) at 1/1000 dilution

    Lane 1 : Ramos cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 44 kDa

  • Flow Cytometry (Intracellular) - Anti-ATG4B antibody [EPR6436(2)] (ab154843)
    Flow Cytometry (Intracellular) - Anti-ATG4B antibody [EPR6436(2)] (ab154843)

    Intracellular Flow Cytometry analysis of permeabilized Jurkat cells labeling ATG4B (red), using ab154843 at a 1/100 dilution, and negative control cells probed with a Rabbit IgG (green)

     

  • Anti-ATG4B antibody [EPR6436(2)] (ab154843)
    Anti-ATG4B antibody [EPR6436(2)] (ab154843)

References (5)

Publishing research using ab154843? Please let us know so that we can cite the reference in this datasheet.

ab154843 has been referenced in 5 publications.

  • Cantarero L  et al. Mitochondria-lysosome membrane contacts are defective in GDAP1-related Charcot-Marie-Tooth disease. Hum Mol Genet 29:3589-3605 (2021). PubMed: 33372681
  • Wang Y  et al. S100A4 Silencing Facilitates Corneal Wound Healing After Alkali Burns by Promoting Autophagy via Blocking the PI3K/Akt/mTOR Signaling Pathway. Invest Ophthalmol Vis Sci 61:19 (2020). PubMed: 32926102
  • Li Z  et al. Targeting the miR-665-3p-ATG4B-autophagy axis relieves inflammation and apoptosis in intestinal ischemia/reperfusion. Cell Death Dis 9:483 (2018). PubMed: 29706629
  • Li L  et al. Forkhead box O3 (FoxO3) regulates kidney tubular autophagy following urinary tract obstruction. J Biol Chem 292:13774-13783 (2017). PubMed: 28705935
  • Russ DW  et al. Muscle-specificity of age-related changes in markers of autophagy and sphingolipid metabolism. Biogerontology 16:747-59 (2015). WB ; Rat . PubMed: 26296420

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-ATG4B antibody [EPR6436(2)]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Fibroblast)
Gel Running Conditions
Reduced Denaturing (4-12)
Loading amount
20 µg
Treatment
knockdown
Specification
Fibroblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Jun 16 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATG4B antibody [EPR6436(2)]

 Excellent
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Culture fibroblast)
Permeabilization
No
Specification
Culture fibroblast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Apr 02 2021

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