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Cell Biology Apoptosis Nucleus PARP
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Apoptosis and DNA Damage (H2A.X(S139) + cleaved PARP1 + Anti-GAPDH) Western Blot Cocktail (ab131385)

  • Datasheet
  • SDS
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Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
  • Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
  • Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)

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Overview

  • Product name

    Apoptosis and DNA Damage (H2A.X(S139) + cleaved PARP1 + Anti-GAPDH) Western Blot Cocktail
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    The Apoptosis and DNA Damage (H2A.X(S139) + cleaved PARP1 + Anti-GAPDH) Western Blot Cocktail (ab131385) is designed to study the induction of DNA damage and/or apoptosis in response to various stimuli. The two main components of this cocktail are monoclonal antibodies specific to cleaved-PARP1 and H2A.X phospho Ser139. H2A.X is a histone H2A family member that is phosphorylated and recruited to sites of double-strand DNA breaks. Poly [ADP-ribose] polymerase 1 (PARP1) is a DNA repair enzyme that is cleaved by activated caspases. Combined, these antibodies provide biomarkers of dsDNA breaks (H2A.X phospho Ser139) and apoptosis (cleaved-PARP1). An anti-GAPDH antibody is included as a loading control. These three readouts are easily resolved by western blot given their different molecular weights.

  • Notes

    Individual antibodies within the ab131385 cocktail:

    Mouse phospho-H2A.X (pSer139) [9F3] monoclonal, IgG
    Working concentration: 1 µg/ml

    Mouse cleaved-PARP1 [4B5BD2] monoclonal, IgG1
    Working concentration: 1 µg/ml 

    Mouse GAPDH [3E8AD9] monoclonal, IgG2b:
    Working concentration: 0.1 µg/ml

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 200 µl
    Antibody cocktail 1 x 100µg
  • Research areas

    • Cell Biology
    • Apoptosis
    • Nucleus
    • PARP
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Huntington's disease
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Epigenetics and Nuclear Signaling
    • Histones
    • Variants
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • ADP-ribosylation
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • Base Excision Repair
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • DNA Damage Recognition
    • Epigenetics and Nuclear Signaling
    • Histones
    • H2A
    • Unmodified
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Epigenetics and Nuclear Signaling
    • Histones
    • H2A
    • Phosphorylated
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Apoptosis
    • Nuclear
    • Epigenetics and Nuclear Signaling
    • Histones
    • H2A
    • Variants
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • PARP
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Kits/ Lysates/ Other
    • Kits
    • Antibody Cocktails
    • Apoptosis & Cell Cycle
  • Cellular localization

    Cleaved PARP1: Nucleus. Nucleus, nucleolus. Localizes at sites of DNA damage. Histone H2A.X: Nucleus. Chromosome. GAPDH: Cytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
  • Database links

    • Entrez Gene: 142 Human
    • Entrez Gene: 2597 Human
    • Entrez Gene: 3014 Human
    • Omim: 138400 Human
    • Omim: 173870 Human
    • Omim: 601772 Human
    • SwissProt: P04406 Human
    • SwissProt: P09874 Human
    • SwissProt: P16104 Human
    • Unigene: 177766 Human
    • Unigene: 477879 Human
    • Unigene: 544577 Human
    • Unigene: 592355 Human
    • Unigene: 598320 Human
    see all

Associated products

  • Related Products

    • Staurosporine, Protein kinase inhibitor (ab120056)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab131385 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
1/250. Predicted molecular weight: 15, 36, 89 kDa. (Entire WB procedure to be done in 4% milk/PBS. Note that pH2A.X is ~16 kDa, hence do not run the SDS-PAGE gel too long. Use anti-Mouse-HRP secondary antibody.)
Notes
WB
1/250. Predicted molecular weight: 15, 36, 89 kDa. (Entire WB procedure to be done in 4% milk/PBS. Note that pH2A.X is ~16 kDa, hence do not run the SDS-PAGE gel too long. Use anti-Mouse-HRP secondary antibody.)

Images

  • Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
    Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
    All lanes :
    Top panel: H2A.X (pSer139) antibody at 1 µg/mL.

    Bottom panel: total H2A.X antibody at a 1/2000 dilution


    Lane 1 : Jurkat cell lysate, untreated
    Lane 2 : Jurkat cell lysate, 4h post UV exposure
    Lane 3 : Jurkat cell lysate, 4h post UV exposure, treated with phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Top panel: Goat anti Mouse HRP at a 1/3000 dilution

    Bottom panel: Goat anti Rat HRP at a 1/3000 dilution

    Predicted band size: 15, 36, 89 kDa

  • Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
    Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
    All lanes : Apoptosis and DNA Damage (H2A.X(S139) + cleaved PARP1 + Anti-GAPDH) Western Blot Cocktail (ab131385) at 1/250 dilution (DNA Damage and Apoptosis cocktail (ab131384) )

    Lane 1 : HeLa cell lysate, untreated
    Lane 2 : HeLa cell lysate, 20 µM Camptothecin 4h treatment
    Lane 3 : HeLa cell lysate, 1 µM Staurosporin 4h treatment

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat anti Mouse HRP at 1/3000 dilution

    Predicted band size: 15, 36, 89 kDa

  • Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
    Western blot - Anti-GAPDH + H2A.X(S139) + cleaved PARP1 antibody (ab131385)
    All lanes : Apoptosis and DNA Damage (H2A.X(S139) + cleaved PARP1 + Anti-GAPDH) Western Blot Cocktail (ab131385) at 1/250 dilution (Damage and Apoptosis cocktail (ab131384) )

    Lane 1 : Jurkat cell lysate, untreated
    Lane 2 : Jurkat cell lysate, 1h post UV exposure
    Lane 3 : Jurkat cell lysate, 2h post UV exposure
    Lane 4 : Jurkat cell lysate, 4h post UV exposure

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti Mouse HRP at 1/3000 dilution

    Predicted band size: 15, 36, 89 kDa

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab131385? Please let us know so that we can cite the reference in this datasheet.

ab131385 has been referenced in 2 publications.

  • Yu Z  et al. ATPase copper transporter A, negatively regulated by miR-148a-3p, contributes to cisplatin resistance in breast cancer cells. Clin Transl Med 10:57-73 (2020). PubMed: 32508020
  • McMahon KA  et al. Identification of intracellular cavin target proteins reveals cavin-PP1alpha interactions regulate apoptosis. Nat Commun 10:3279 (2019). PubMed: 31332168

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