Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20ºC. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR6436(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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 |
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Flow Cyt (Intra) |
1/100 - 1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 44 kDa.
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Notes |
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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. |
Target
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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
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Database links
- Entrez Gene: 23192 Human
- Entrez Gene: 66615 Mouse
- Entrez Gene: 316640 Rat
- Omim: 611338 Human
- SwissProt: Q9Y4P1 Human
- SwissProt: Q8BGE6 Mouse
- SwissProt: Q62625 Rat
- Unigene: 283610 Human
see all -
Alternative names
- APG4 autophagy 4 homolog B antibody
- APG4B antibody
- ATG4 autophagy related 4 homolog B (S. cerevisiae) antibody
see all
Images
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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.
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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.
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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 -
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)
Datasheets and documents
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SDS download
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Datasheet download
References (6)
ab154843 has been referenced in 6 publications.
- Zhang Q et al. BECLIN-1-Mediated Autophagy Suppresses Silica Nanoparticle-Induced Testicular Toxicity via the Inhibition of Caspase 8-Mediated Cell Apoptosis in Leydig Cells. Cells 11:N/A (2022). PubMed: 35740992
- 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