Overview

  • Product name
  • Description
    Rabbit polyclonal to Endo G
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ELISA, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    GGPRGPGELAKYGLP

    , corresponding to amino acids 55-70 of Human Endo G. (Peptide available as ab9648.)

  • Positive control
    • HepG2 and 3T3 cell lysates
  • General notes


    The fragmentation of nuclear DNA is a hallmark of apoptotic cell death. The activities of caspase and nuclease are involved in the DNA fragmentation. Caspase-activated deoxyribonuclease (CAD), also termed DNA fragmentation factor (DFF40), is one such nuclease, and is capable of inducing DNA fragmentation and chromatin condensation after cleavage by caspase-3 of its inhibitor ICAD/DFF45. Caspase and CAD independent DNA fragmentation also exists. Recent studies demonstrated that another nuclease, endonuclease G (endoG), is specifically activated by apoptotic stimuli and is able to induce nucleosomal fragmentation of DNA independently of caspase and DFF/CAD (1,2). EndoG is a mitochondrion-specific nuclease that translocates to the nucleus and cleaves chromatin DNA during apoptosis. The homologue of mammalian EndoG is the first mitochondrial protein identified to be involved in apoptosis in C. elegans (2). EndooG also cleaves DNA in vitro (4).

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Primary antibody notes
    The fragmentation of nuclear DNA is a hallmark of apoptotic cell death. The activities of caspase and nuclease are involved in the DNA fragmentation. Caspase-activated deoxyribonuclease (CAD), also termed DNA fragmentation factor (DFF40), is one such nuclease, and is capable of inducing DNA fragmentation and chromatin condensation after cleavage by caspase-3 of its inhibitor ICAD/DFF45. Caspase and CAD independent DNA fragmentation also exists. Recent studies demonstrated that another nuclease, endonuclease G (endoG), is specifically activated by apoptotic stimuli and is able to induce nucleosomal fragmentation of DNA independently of caspase and DFF/CAD (1,2). EndoG is a mitochondrion-specific nuclease that translocates to the nucleus and cleaves chromatin DNA during apoptosis. The homologue of mammalian EndoG is the first mitochondrial protein identified to be involved in apoptosis in C. elegans (2). EndooG also cleaves DNA in vitro (4).
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9647 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
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 35 kDa.Can be blocked with Human Endo G peptide (ab9648).
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Relevance
    Endo G is a nuclear encoded endonuclease that is localized in the mitochondrion. The encoded protein cleaves DNA at GC tracts. It is capable of generating the RNA primers required by DNA polymerase gamma to initiate replication of mitochondrial DNA.
  • Cellular localization
    Cytoplasmic, Mitochondrial and Nuclear
  • Database links
  • Alternative names
    • EndoG antibody
    • Endonuclease G antibody
    • Endonuclease G mitochondrial antibody
    • EndonucleaseG antibody
    • FLJ27463 antibody
    • Mitochondrial endonuclease G antibody
    • NUCG_HUMAN antibody
    see all

Images

  • Western blot analysis of EndoG in mouse (M) 3T3 and human (H) HepG2 cell lysates with anti-EndoG at 2 µg/ml.

    Western blot analysis of EndoG in mouse (M) 3T3 and human (H) HepG2 cell lysates with anti-EndoG at 2 µg/ml.
  • ab9647 at 1/50 dilution staining Endo G in human pancreas by Immunohistochemistry (Formalin/ PFA fixed paraffin embedded sections).

  • ICC/IF image of ab9647 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9647, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Peeva V  et al. Linear mitochondrial DNA is rapidly degraded by components of the replication machinery. Nat Commun 9:1727 (2018). Read more (PubMed: 29712893) »
  • Chen S  et al. Alpha lipoic acid attenuates cadmium-induced nephrotoxicity via the mitochondrial apoptotic pathways in rat. J Inorg Biochem 184:19-26 (2018). Read more (PubMed: 29654931) »
See all 22 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skeletal muscle)
Permeabilization
Yes - 0.5% Triton
Specification
Skeletal muscle
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 20 2017

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12% Gel)
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 04 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12% Gel)
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 04 2014

Question
Answer

Merci de votre intérêt pour ab9647. Comme discuté au téléphone, cet anticorps n’a pas encore été testé en cytométrie de flux . Cependant, si vous souhaitez tester ab9647 en cytométrie de flux, il me sera possible de vous offrir une remise. Après soumission d’une Abreview avec vos résultats, une remise correspondant à la valeur d’achat de ab9647 pourra être utilisable lors d’une prochaine commande d’anticorps primaire. Etapes à suivre : 1. Nous confirmer que vous souhaiteriez tester ab9647 en cytométrie de flux afin de recevoir le code de remise correspondant. Ce code doit être édité avant l’achat de ab9647 2. Acheter ab9647 par téléphone, fax ou internet (www.abcam.com) 3. Le tester en cytométrie de flux 4. Nous faire part de vos résultats, positifs ou négatifs, grâce à notre système Abreview. Pour plus d’informations https://www.abcam.com/abreviews 5. Après soumission de votre Abreview, appeler notre service clientèle afin de passer votre prochaine commande grâce au code de réduction. Ce code de réduction est utilisable 120 jours après son édition et utilisable lors de l’achat d’un autre anticorps primaire. Même si ab9647 est démontré ne pas fonctionner en cytométrie de flux, après soumission de votre Abreview, vous recevrez quand même une remise sur l’achat de votre prochain anticorps. N’hésitez pas à nous demander plus d’informations concernant cette offre promotionnelle. Termes et Conditions : www.abcam.com/collaborationdiscount.

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Answer

Please refer to the "Protocols" section (IHC, Western blot, ELISA, FACS etc) on the right hand side panel of the Abcam homepage. You may find some useful information there.

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Question
Answer

All the information available for this product is listed on the on-line datasheet (price, datasheet, publication, suitability, cross-reactivity). According to the datasheet, this antibody has not been tested for IP application.

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Answer

We have never tested this antibody in IHC, therefore we can't guarantee that it will work. To the best of our knowledge, no-one else has tested it in IHC, either.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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