ATG3 Recombinant Monoclonal Antibody

Datasheet

Code	CSB-RA794271A0HU
Size	US$210
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Image	Western Blot Positive WB detected in: Hela whole cell lysate(30µg), A431 whole cell lysate(30µg), MCF-7 whole cell lysate(30µg), K562 whole cell lysate(30µg), PC-12 whole cell lysate(30µg), THP-1 whole cell lysate(30µg), U-87MG whole cell lysate(30µg), Mouse brain tissue lysate(30µg) All lanes: ATG3 antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 36 kDa Observed band size: 40 kDa Exposure time：2min IHC image of CSB-RA794271A0HU diluted at 1:100 and staining in paraffin-embedded human small intestine tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. IHC image of CSB-RA794271A0HU diluted at 1:100 and staining in paraffin-embedded human colorectal cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Overlay Peak curve showing Hela cells stained with CSB-RA794271A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10, 000 events was performed.
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Target Background

Product Details

Uniprot No.

Q9NT62

Target Names

ATG3

Alternative Names

2610016C12Rik antibody; Apg 3 antibody; APG3 antibody; APG3 autophagy 3 like antibody; APG3 like antibody; APG3, S. cerevisiae, homolog of antibody; APG3-like antibody; APG3L antibody; Apg3p antibody; ATG 3 antibody; ATG3 antibody; ATG3 autophagy related 3 homolog antibody; ATG3 autophagy related 3 homolog (S. cerevisiae) antibody; ATG3_HUMAN antibody; Autophagy 3, S. cerevisiae, homolog of antibody; Autophagy Apg3p/Aut1p like antibody; autophagy related 3 antibody; Autophagy related protein 3 antibody; Autophagy-related protein 3 antibody; DKFZp564M1178 antibody; FLJ22125 antibody; hApg3 antibody; MGC15201 antibody; OTTHUMP00000214547 antibody; OTTHUMP00000214548 antibody; PC3 96 antibody; Protein PC3-96 antibody; Ubiquitin-like-conjugating enzyme ATG3 antibody

Species Reactivity

Human, Mouse

Immunogen

A synthesized peptide from human ATG3 protein

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

3F10

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
IHC	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

ATG3 serves as a critical E2-like conjugating enzyme in the autophagy pathway, facilitating the lipidation of LC3 proteins essential for autophagosome formation. This process plays a fundamental role in cellular homeostasis, stress responses, and has significant implications in cancer biology, neurodegeneration, and metabolic disorders, making ATG3 a valuable target for researchers investigating autophagy mechanisms.

This recombinant monoclonal antibody, developed using rabbit host immunization against a synthetic peptide derived from human ATG3, offers the reproducibility and consistency that demanding autophagy research requires. As a sequence-defined recombinant clone, it eliminates the lot-to-lot variability that can complicate longitudinal studies, ensuring your experimental conditions remain constant across projects.

Validation data demonstrates robust performance across multiple applications. In Western blot analysis, the antibody detects ATG3 at approximately 40 kDa across a diverse panel of human cell lines including HeLa, A431, MCF-7, K562, THP-1, and U-87MG, as well as mouse brain tissue lysate, confirming cross-species reactivity with both human and mouse samples. The apparent molecular weight of the band exceeds the predicted 36 kDa. This may be explained by either the presence of low-abundance post-translational modifications (such as glycosylation or phosphorylation) on ATG3 under physiological conditions, or by the deviation in SDS-PAGE mobility caused by its amino acid composition. Immunohistochemistry validation in paraffin-embedded human small intestine and colorectal cancer tissues demonstrates clear staining patterns suitable for tissue-based studies. Flow cytometry analysis in HeLa cells shows distinct signal separation from isotype controls, supporting intracellular detection applications.

Whether you are investigating autophagy flux, screening therapeutic compounds targeting this pathway, or characterizing ATG3 expression in disease models, this antibody provides the flexibility to move seamlessly between Western blot, immunohistochemistry, and flow cytometry workflows within your cell biology research.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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Target Background

Function

E2 conjugating enzyme required for the cytoplasm to vacuole transport (Cvt), autophagy, and mitochondrial homeostasis. Responsible for the E2-like covalent binding of phosphatidylethanolamine to the C-terminal Gly of ATG8-like proteins (GABARAP, GABARAPL1, GABARAPL2 or MAP1LC3A). The ATG12-ATG5 conjugate plays a role of an E3 and promotes the transfer of ATG8-like proteins from ATG3 to phosphatidylethanolamine (PE). This step is required for the membrane association of ATG8-like proteins. The formation of the ATG8-phosphatidylethanolamine conjugates is essential for autophagy and for the cytoplasm to vacuole transport (Cvt). 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. Promotes primary ciliogenesis by removing OFD1 from centriolar satellites via the autophagic pathway.

Gene References into Functions

ATG3 was targeted by miR-23a, suggesting that ATG3 is an autophagy-related competing endogenous RNA. PMID: 30007957
Results revealed that miR-16 was significantly downregulated, and ATG3 was significantly upregulated in non-small cell lung carcinoma (NSCLC) patient tissue samples. ATG3 was found to be a direct target of miR-16. PMID: 29138833
miR-155 silencing rescued autophagosome number in Mycobacterium tuberculosis infected dendritic cells and enhanced autolysosome fusion, thereby supporting a previously unidentified role of the miR-155 as inhibitor of ATG3 expression. PMID: 29300789
PTK2 inhibition-induced sustained levels of ATG3 were able to sensitize cancer cells to DNA-damaging agents. PMID: 28103122
Our data demonstrate that HOTAIR promotes the activation of autophagy in HCC cells by upregulating the expression of the autophagy-related genes ATG3 and ATG7. PMID: 27301338
ATG3 upregulation contributes to autophagy induced by the detachment of intestinal epithelial cells from the extracellular matrix, but promotes autophagy-independent apoptosis of the attached cells PMID: 26061804
Hidden Markov models were used to detect protein homology among the flexible regions of Atg3 homologs and importance of conserved regions evaluated by performing affinity capture experiments with human Atg3 deletion constructs; binding studies and competition experiments demonstrate that overlapping sites in the Atg3FR are important for E3 binding and E1 binding. PMID: 24186333
The region of human ATG3 that interacts with ATG7 is precisely identified using nuclear magnetic resonance. PMID: 26043688
ATG3 gene and its gene family may play an important role in transformation of myelodysplastic syndrome. PMID: 24420857
Lipidation of the LC3/GABARAP family of autophagy proteins relies on a membrane-curvature-sensing domain in Atg3. PMID: 24747438
13 residues of the ATG3 fragment form a short beta-strand followed by an alpha-helix on a surface area that is an exclusive binding site for ATG12. PMID: 24191030
caspase-8 overexpression led to Atg3 degradation and this event depended on caspase-8 enzymatic activity PMID: 22644571
These results unveil a role for ATG12-ATG3 in mitochondrial homeostasis and implicate the ATG12 conjugation system in cellular functions distinct from the early steps of autophagosome formation. PMID: 20723759
Human Apg3p/Aut1p homologue is an authentic E2 enzyme for multiple substrates, GATE-16, GABARAP, and MAP-LC3, and facilitates the conjugation of hApg12p to hApg5p PMID: 11825910
Murine Atg8L/Apg8L modification is mediated by human Atg3. PMID: 16704426

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Subcellular Location

Cytoplasm.

Protein Families

ATG3 family

Tissue Specificity

Widely expressed, with a highest expression in heart, skeletal muscle, kidney, liver and placenta.

Database Links

HGNC: 20962

OMIM: 609606

KEGG: hsa:64422

STRING: 9606.ENSP00000283290

UniGene: Hs.477126

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Antibody FAQs What Is an Antibody? How to Choose an Antibody for Scientific Research? The difference between primary antibody and secondary antibod The Meaning of Irregular Antibody Screening Tag Antibodies, Your Good Partner in Protein Research How to Choose the Loading Control Antibodies?

Related pathways

Autophagy

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301-363-4651 (Available 9 a.m. to 5 p.m. CST from Monday to Friday)

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Address

No. 269, Shendun 5th Road, Donghu Hi-Tech Development Area, Hubei Province, 430206, P.R.China

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