SpecificityIHC, WB and ELISA confirmed the specificity for ATG3. A 37 kDa band, that corresponds to the molecular weight of ATG3, is detected via western blot analysis. Human, rat, mouse. Other species not yet tested.
Species ReactivityHuman, Mouse, Rat
Immunogen DescriptionA synthetic peptide corresponding to a central region of the human ATG3 protein [aa. 151 EGEAADMEEYE 161] conjugated to Blue Carrier Protein has been used as the immunogen. The peptide is highly conserved and homologous with the corresponding sequence derived from ATG3 protein in mouse, rat, S. cerevisiae, Macaca mulatta (monkey), Canis familiaris (dog), and many more species.
Application DetailsIHC, WB, immunofluorescence. A dilution of 1:50 to 1:3000 dilution is recommended for these applications. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetAutophagy-related protein 3 (ATG3)
SpecificityIHC, WB and ELISA confirmed the specificity for ATG3. A 37 kDa band, that corresponds to the molecular weight of ATG3, is detected via western blot analysis. Human, rat, mouse. Other species not yet tested.
Target Host SpeciesHuman
Species ReactivityHuman, Mouse, Rat
Antibody HostRabbit
Antibody TypePolyclonal
Antibody IsotypeMixed
ConjugateUnconjugated
Immunogen DescriptionA synthetic peptide corresponding to a central region of the human ATG3 protein [aa. 151 EGEAADMEEYE 161] conjugated to Blue Carrier Protein has been used as the immunogen. The peptide is highly conserved and homologous with the corresponding sequence derived from ATG3 protein in mouse, rat, S. cerevisiae, Macaca mulatta (monkey), Canis familiaris (dog), and many more species.
Immunogen Length11 amino acids
SequenceEGEAADMEEYE
EpitopeAmino acids 151-161 of human ATG3 protein
Purity DescriptionWhole serum
FormatLyophilized
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution keep aliquots at -20°C for a higher stability, and at 2-8°C with an appropriate antibacterial agent. Glycerol (1:1) may be added for an additional stability. Avoid repetitive freeze/thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesAutophagy-related protein 3; APG3; APG3-like; hApg3; Protein PC3-96 ; ATG3; APG3; APG3L
Scientific BackgroundFUNCTION: GABARAPL1 (GABARAPL2 or GABARAP or MAP1LC3)-modifier protein conjugating enzyme involved in its E2-like covalent binding to PE. ATG7 (E1-like enzyme) facilitates this reaction by forming an E1-E2 complex with ATG3 (E2-like enzyme). Preferred substrate is MAP1LC3A. Formation of the GABARAPL1-PE conjugate is essential for autophagy. SUBUNIT: Interacts with ATG7 and ATG12. The complex, composed of ATG3 and ATG7, plays a role in the conjugation of ATG12 to ATG5. SUBCELLULAR LOCATION: Cytoplasm. ALTERNATIVE PRODUCTS: 2 named isoforms produced by alternative splicing. TISSUE SPECIFICITY: Widely expressed, with a highest expression in heart, skeletal muscle, kidney, liver and placenta. SIMILARITY: Belongs to the ATG3 family.
Confocal microscopy on immunofluorescently detected ATG3 in cytospin-isolated human white blood cells using Rabbit antibody to ATG3: whole serum (R-159-100) at a dilution of 1: 100, incubated for 1 h at room temperature. The cells stained for ATG3 appear in red . The cells were counter stained with Hoechst Dye (blue colour). Here, the merged picture is presented. More information on the staining: Fixation: In acetone for 90 seconds. Antibody dilutions: Primary antibody was diluted 1:100 and incubated for 1 hour, washed then 3x, and the secondary (Goat anti Rb 1:100 Alexa 546 conjugate) was incubated for 30 minutes. Magnification 80X. No staining was observed using the Pre-immunisation serum. No staining was observed in HL60 cell line using Rabbit antibody to ATG3: whole serum (R-159-100)
Western blot on mouse brain lysate using Rabbit antibody to ATG3: whole serum (R-159-100) at a dilution of 1: 50. Absorbption with the immunising peptide almost removed the three main bands otherwise visible on the unabsorbed strip.
Western blot under reducing conditions on 293 cell lysate using Rabbit antibody to ATG3: whole serum (R-159-100) at a dilution of 1: 50.
Western blot under reducing conditions on Odora (an Olfactory cell line) cell lysate using Rabbit antibody to ATG3: whole serum (R-159-100) at a dilution of 1: 50.
Specific ReferencesRyningen A et al (2012) Inhibition of Mammalian target of rapamycin in human acute myeloid leukemia cells has diverse effects that depend on the environmental in vitro stress. Bone Marrow Res. 2012;2012:329061.
General ReferencesTanida I, et al. J. Biol. Chem. 277:13739-13744(2002). Ota T, et al. Nat. Genet. 36:40-45(2004). Rush J, et al. Nat. Biotechnol. 23:94-101(2005).