Name: Anti-ATG3 antibody [EPR4801]
Brand: RabMAb
SKU: ab108251
Rating: 4 (6 reviews)

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Primary - Rabbit Anti-ATG3 antibody [EPR4801] (ab108251)
IHC-Fr, WB, IHC-P, Flow Cyt, ICC/IF

Protein - Recombinant Human ATG3 protein (ab152071)
SDS-PAGE, HPLC

View more associated products

Product name

Anti-ATG3 antibody [EPR4801]
See all ATG3 primary antibodies
Description

Rabbit monoclonal [EPR4801] to ATG3
Host species

Rabbit
Tested applications

Suitable for: IHC-Fr, WB, IHC-P, Flow Cyt, ICC/IFmore details
Unsuitable for: IP
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide within Human ATG3 (N terminal). The exact sequence is proprietary.
Positive control
- WB: K562, HL-60, HeLa or Jurkat whole cell lysate (ab7899). IHC-P: Human muscle tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR4801
Isotype

IgG
Research areas

Alternative Versions
- Anti-ATG3 antibody [EPR4801] - BSA and Azide free (ab226019)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- Jurkat cytoplasmic extract lysate (ab14842)
Recombinant Protein
- Recombinant Human ATG3 protein (ab152071)

Our Abpromise guarantee covers the use of ab108251 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
IHC-Fr		Use at an assay dependent concentration.
WB		1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Perform antigen retrieval before commencing with IHC staining protocol.
Flow Cyt		1/2000.
ICC/IF		1/100 - 1/250.

Application notes

Is unsuitable for IP.

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.
Target information above from: UniProt accession Q9NT62 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 64422 Human
- Entrez Gene: 67841 Mouse
- Entrez Gene: 171415 Rat
- Omim: 609606 Human
- SwissProt: Q9NT62 Human
- SwissProt: Q9CPX6 Mouse
- SwissProt: Q6AZ50 Rat
- Unigene: 477126 Human
- Unigene: 41775 Mouse
- Unigene: 3084 Rat
see all
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
see all

Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)

All lanes : Anti-ATG3 antibody [EPR4801] (ab108251) at 1/10000 dilution

Lane 1 : K562 cell lysate
Lane 2 : HL-60 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit

Predicted band size: 36 kDa
Observed band size: 40 kDa
why is the actual band size different from the predicted?
Immunohistochemistry (Frozen sections) - Anti-ATG3 antibody [EPR4801] (ab108251)This image is courtesy of an anonymous Abreview

ab108251 staining ATG3 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.
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody [EPR4801] (ab108251)

Immunofluorescent staining of HeLa cells using ab108251 at 1/100.
Flow Cytometry - Anti-ATG3 antibody [EPR4801] (ab108251)

Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATG3 (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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG3 antibody [EPR4801] (ab108251)

ab108251, at a 1/100 dilution, staining Human ATG3 in muscle, using Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Datasheet

This product has been referenced in:

Qiu S et al. The blockade of lipophagy pathway is necessary for docosahexaenoic acid to regulate lipid droplet turnover in hepatic stellate cells. Biomed Pharmacother 109:1841-1850 (2019). Read more (PubMed: 30551439) »
Agrotis A et al. Redundancy of human ATG4 protease isoforms in autophagy and LC3/GABARAP processing revealed in cells. Autophagy 15:976-997 (2019). Read more (PubMed: 30661429) »

See all 8 Publications for this product

Publishing research using ab108251? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Rat Cell (H9c2 rat cardiomyocytes)

Permeabilization

Yes - 0.01% TritonX-100

Specification

H9c2 rat cardiomyocytes

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 18 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (Cardiomyocytes)

Permeabilization

Yes - 0.01% TritonX-100

Specification

Cardiomyocytes

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 18 2020

Application

Western blot

Sample

Mouse Tissue lysate - whole (Lung)

Gel Running Conditions

Reduced Denaturing (4-20%)

Loading amount

15 µg

Treatment

1x10^7 S. aureus peritoneal clot

Specification

Lung

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Mr. Alan Chang

Verified customer

Submitted Mar 20 2014

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (uterus)

Permeabilization

Specification

uterus

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Formaldehyde

Dr. Varkha Agrawal

Verified customer

Submitted Nov 14 2013

Application

Western blot

Sample

Mouse Tissue lysate - whole (placenta)

Gel Running Conditions

Reduced Denaturing (4-12)

Loading amount

5 µg

Specification

placenta

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Varkha Agrawal

Verified customer

Submitted Nov 14 2013

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (Uterus)

Permeabilization

Specification

Uterus

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C

Fixative

Paraformaldehyde

Dr. Mukesh Jaiswal

Verified customer

Submitted Nov 13 2013

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

Positive Controls

Recombinant Protein

Applications

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATG3 antibody [EPR4801]

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATG3 antibody [EPR4801]

Western blot abreview for Anti-ATG3 antibody [EPR4801]

Immunohistochemistry (Frozen sections) abreview for Anti-ATG3 antibody [EPR4801]

Western blot abreview for Anti-ATG3 antibody [EPR4801]

Immunohistochemistry (Frozen sections) abreview for Anti-ATG3 antibody [EPR4801]

Post-translational
modifications