Product datasheet
Anti-Apg3 antibody ab66287
2 Images
Overview
Product name
Anti-Apg3 antibody
Description
Rabbit polyclonal to Apg3
Tested applications
ICC/IF, WB
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Sheep, Cow, Dog
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human
Apg3.Read Abcam's proprietary immunogen policy(Peptide available as ab66286.)
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)
Properties
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
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
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).
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Target
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
modifications
Conjugated 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.
Anti-Apg3 antibody images
All lanes : Anti-Apg3 antibody (ab66287) at
1 µg/ml
Lane 1 : TE 671 (Human
Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 : Raji (Human Burkitt's lymphoma cell
line) Whole Cell Lysate - tumor cell line
(ab30124)
Lane 3 : JEG-3 (Human placental
choriocarcinoma cell line) Whole Cell Lysate
Western blot - Anti-Apg3 antibody (ab66287)
Lane 4 : K562 (Human erythromyeloblastoid
leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - PreAdsorbed (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.
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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 nonspecific protein-protein interactions. The cells
Immunocytochemistry/ Immunofluorescence Apg3 antibody (ab66287)
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.
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