The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).
1/100 - 1/250. Perform antigen retrieval before commencing with IHC staining protocol.
1/100 - 1/250.
Application notesIs unsuitable for IP.
FunctionE2-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 specificityWidely expressed, with a highest expression in heart, skeletal muscle, kidney, liver and placenta.
Sequence similaritiesBelongs 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.
Immunohistochemistry (Frozen sections) - Anti-Apg3 antibody [EPR4801] (ab108251)This image is courtesy of an anonymous Abreview
ab108251 staining Apg3 in Mouse uterus tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with 5% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/200 in 1% BSA in PBS) for 12 hours at 4°C. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.
Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Apg3 (red) with ab108251 at a 1/2000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
ab108251, at a 1/100 dilution, staining Human Apg3 in muscle, using Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue.
References for Anti-Apg3 antibody [EPR4801] (ab108251)
This product has been referenced in:
Russ DW et al. Muscle-specificity of age-related changes in markers of autophagy and sphingolipid metabolism. Biogerontology16:747-59 (2015).
Read more (PubMed: 26296420) »
Haldar AK et al. The E2-like conjugation enzyme Atg3 promotes binding of IRG and Gbp proteins to Chlamydia- and Toxoplasma-containing vacuoles and host resistance. PLoS One9:e86684 (2014).
Read more (PubMed: 24466199) »