Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)

Rabbit Recombinant Monoclonal ATG12 antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF, IP and reacts with Human, Mouse, Rat samples.

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Images

Immunoprecipitation - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (AB303489), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	ICC/IF	Flow Cyt (Intra)	IP
Human	Tested	Tested	Not recommended	Not recommended	Tested
Mouse	Tested	Tested	Tested	Not recommended	Tested
Rat	Tested	Tested	Expected	Not recommended	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Target data

See full target information ATG12

Function

Ubiquitin-like protein involved in autophagy vesicles formation. Conjugation with ATG5 through a ubiquitin-like conjugating system involving also ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme, is essential for its function. The ATG12-ATG5 conjugate acts as an E3-like enzyme which is required for lipidation of ATG8 family proteins and their association to the vesicle membranes. As part of the ATG8 conjugation system with ATG5 and ATG16L1, required for recruitment of LRRK2 to stressed lysosomes and induction of LRRK2 kinase activity in response to lysosomal stress (By similarity). (Microbial infection) May act as a proviral factor. In association with ATG5, negatively regulates the innate antiviral immune response by impairing the type I IFN production pathway upon vesicular stomatitis virus (VSV) infection (PubMed:17709747). Required for the translation of incoming hepatitis C virus (HCV) RNA and, thereby, for the initiation of HCV replication, but not required once infection is established (PubMed:19666601).

Alternative names

APG12, APG12L, ATG12, Ubiquitin-like protein ATG12, Autophagy-related protein 12, APG12-like

Recommended products

Rabbit Recombinant Monoclonal ATG12 antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF, IP and reacts with Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR27485-89
Purification technique: Affinity purification Protein A
Specificity: Unsuitable for human ICC
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

ATG12 also known as Autophagy-related protein 12 plays a critical role in autophagy an essential cellular process for degrading and recycling cellular components. The molecular weight of ATG12 is around 14 kDa. It is part of the ATG12-conjugation system and is invariably expressed in various tissues with notable presence in the liver brain and heart. ATG12 forms a covalent bond with ATG5 in a ubiquitin-like conjugation pathway which is significant for this mechanism.

Biological function summary

ATG12 is vital for the formation of autophagosomes which are double-membraned vesicles that sequester cellular components for degradation. It functions as part of a complex by binding to ATG5 and further associates with ATG16L1 to form a larger ATG12-ATG5-ATG16 complex. This complex localizes to the outer membrane of the growing autophagosome facilitating elongation and closure of the vesicle making it effective in the regulation of autophagy.

Pathways

ATG12's interaction with other proteins like ATG5 places it in the macroautophagy pathway an important process maintaining cellular homeostasis and turnover of cellular components. In addition to this ATG12 also contributes to cellular responses to stress conditions via this pathway. Furthermore it has connections with signaling pathways involving proteins such as LC3 which marks autophagosomal membranes and interacts with ATG12-conjugated complexes enhancing the autophagic flux.

Associated diseases and disorders

Researchers associate dysregulated expression of ATG12 with neurodegenerative diseases such as Parkinson’s disease where impaired autophagy leads to toxic protein accumulation. Additionally alterations in ATG12 expression have links to cancer as enhanced autophagy in some tumor cells supports survival under metabolic stress. Both disorders demonstrate connections to proteins involved in the autophagy pathway suggesting therapeutic potential in modulating ATG12 activity.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Immunoprecipitation - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

ATG12 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10ug with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10ug
Lane 2: Anti-ATG12 antibody [EPR27485-89] ab303488 IP in NIH/3T3 whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-ATG12 antibody [EPR27485-89] ab303488 in NIH/3T3 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 180 seconds
All lanes: Immunoprecipitation - Anti-ATG12 antibody [EPR27485-89] (Anti-ATG12 antibody [EPR27485-89] ab303488) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10ug
Lane 2: Anti-ATG12 antibody [EPR27485-89] ab303488 IP in NIH/3T3 whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa
Exposure time: 180s
Immunoprecipitation - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

ATG12 was immunoprecipitated from 0.35 mg Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

Lane 1: Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2: abAB303488 IP in Hela whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-ATG12 antibody [EPR27485-89] ab303488 in Hela whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 180 seconds
All lanes: Immunoprecipitation - Anti-ATG12 antibody [EPR27485-89] (Anti-ATG12 antibody [EPR27485-89] ab303488) at 1/1000 dilution
Lane 1: Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2: Anti-ATG12 antibody [EPR27485-89] ab303488 IP in Hela whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa
Exposure time: 180s
Western blot - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
In Western blot, Anti-ATG12 antibody [EPR27485-89] ab303488 was shown to bind specifically to Atg5 bound Atg12 proteins. A band was observed at 52kDa in wild-type HAP1 cell lysate with no signal observed at this size in Atg12 knockout cell line.
Exposure time: 180 seconds
All lanes: Western blot - Anti-ATG12 antibody [EPR27485-89] (Anti-ATG12 antibody [EPR27485-89] ab303488) at 1/1000 dilution
Lane 1: Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell line), whole cell lysate at 20 µg
Lane 2: Atg12 knockout HAP1 whole cell lysate at 20 µg
Lane 3: HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 52 kDa
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling ATG12 with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/100 (4.91 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection). Positive staining on rat testis. The section was incubated with Anti-ATG12 antibody [EPR27485-89] ab303488 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling ATG12 with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/50 (9.82 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image of NIH/3T3 cells treated with Chloroquine (200?M) for 2 hours. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded A Wild-type HAP1 (H tissue labeling ATG12 with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/2000 (0.246 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection). Positive staining on (A) wild-type HAP1 cell pellet, no staining on (B) ATG12 knockout HAP1 cell pellet. The section was incubated with Anti-ATG12 antibody [EPR27485-89] ab303488 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse breast carcinoma tissue labeling ATG12 with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/100 (4.91 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection). Positive staining on mouse breast carcinoma. The section was incubated with Anti-ATG12 antibody [EPR27485-89] ab303488 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling ATG12 with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/100 (4.91 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection). Positive staining on human breast carcinoma. The section was incubated with Anti-ATG12 antibody [EPR27485-89] ab303488 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling ATG12 with Anti-ATG12 antibody [EPR27485-89] ab303488 at 1/100 (4.91 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with Anti-ATG12 antibody [EPR27485-89] ab303488 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
Exposure time: Lane 1-2: 125 seconds
Lane 3-4: 26 seconds
All lanes: Western blot - Anti-ATG12 antibody [EPR27485-89] (Anti-ATG12 antibody [EPR27485-89] ab303488) at 1/1000 dilution
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate ####20000#dilution#Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 20 µg
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 52 kDa
Western blot - Anti-ATG12 antibody [EPR27485-89] - BSA and Azide free (ab303489)
This data was developed using Anti-ATG12 antibody [EPR27485-89] ab303488, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
Exposure time: 26 seconds
All lanes: Western blot - Anti-ATG12 antibody [EPR27485-89] (Anti-ATG12 antibody [EPR27485-89] ab303488) at 1/1000 dilution
Lane 1: Mouse pancreas tissue lysate at 20 µg
Lane 2: Mouse testis tissue lysate at 20 µg
Lanes 3 - 4: Rat brain tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 52 kDa
Exposure time: 26s