Anti-ATG3 antibody (ab66287)
Key features and details
- Rabbit polyclonal to ATG3
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-ATG3 antibody
See all ATG3 primary antibodies -
Description
Rabbit polyclonal to ATG3 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Sheep, Cow, Dog
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Immunogen
Synthetic peptide corresponding to Human ATG3 aa 200-300 conjugated to keyhole limpet haemocyanin.
(Peptide available asab66286) -
Positive control
- This antibody gave a positive signal in the following Human Whole Cell Lysates: TE 671, Raji, JEG-3, K562, HeLa - Staurosporine Treated (48hr, 500nM) (data not shown)
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab66287 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 ICC/IF Use a concentration of 5 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa). Target
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Function
E2-like enzyme involved in autophagy and mitochondrial homeostasis. Catalyzes the conjugation of ATG8-like proteins (GABARAP, GABARAPL1, GABARAPL2 or MAP1LC3A) to phosphatidylethanolamine (PE). PE-conjugation to ATG8-like proteins is essential for autophagy. Preferred substrate is MAP1LC3A. Also acts as an autocatalytic E2-like enzyme, catalyzing the conjugation of ATG12 to itself, ATG12 conjugation to ATG3 playing a role in mitochondrial homeostasis but not in autophagy. ATG7 (E1-like enzyme) facilitates this reaction by forming an E1-E2 complex with ATG3. -
Tissue specificity
Widely expressed, with a highest expression in heart, skeletal muscle, kidney, liver and placenta. -
Sequence similarities
Belongs to the ATG3 family. -
Post-translational
modificationsConjugated to ATG12 at Lys-243. ATG12-conjugation plays a role in regulation of mitochondrial homeostasis and cell death, while it is not involved in PE-conjugation to ATG8-like proteins and autophagy. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 508571 Cow
- Entrez Gene: 64422 Human
- Entrez Gene: 67841 Mouse
- Entrez Gene: 171415 Rat
- Omim: 609606 Human
- SwissProt: Q0VCL3 Cow
- SwissProt: Q9NT62 Human
- SwissProt: Q9CPX6 Mouse
see all -
Alternative names
- 2610016C12Rik antibody
- Apg 3 antibody
- APG3 antibody
see all
Images
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Western blot - Anti-ATG3 antibody (ab66287)All lanes : Anti-ATG3 antibody (ab66287) at 1 µg/ml
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 :Raji whole cell lysate (ab30124)
Lane 3 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate
Lane 4 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 36 kDa
Additional bands at: 72 kDa. We are unsure as to the identity of these extra bands. -
Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody (ab66287)ICC/IF image of ab66287 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66287, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) MCF-7 cells at 5µg/ml.
Protocols
Datasheets and documents
References (1)
ab66287 has been referenced in 1 publication.
- Huang Y et al. Phospho-?Np63a-dependent regulation of autophagic signaling through transcription and micro-RNA modulation. Cell Cycle 11:1247-59 (2012). PubMed: 22356768
Images
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Western blot - Anti-ATG3 antibody (ab66287)All lanes : Anti-ATG3 antibody (ab66287) at 1 µg/ml
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 :Raji whole cell lysate (ab30124)
Lane 3 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate
Lane 4 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 36 kDa
Additional bands at: 72 kDa. We are unsure as to the identity of these extra bands. -
Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody (ab66287)ICC/IF image of ab66287 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66287, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) MCF-7 cells at 5µg/ml.
