Apoptosis and DNA Damage (H2A.X(S139) + cleaved PARP1 + Anti-GAPDH) Western Blot Cocktail (ab131385)
Overview
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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.
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Notes
Individual antibodies within the ab131385 cocktail:
Mouse phospho-H2A.X (pSer139) [9F3] monoclonal, IgG
Working concentration: 1 µg/mlMouse cleaved-PARP1 [4B5BD2] monoclonal, IgG1
Working concentration: 1 µg/mlMouse GAPDH [3E8AD9] monoclonal, IgG2b:
Working concentration: 0.1 µg/ml -
Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 200 µl Antibody cocktail 1 x 100µg -
Research areas
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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
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Associated products
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Related Products
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.)
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| Notes |
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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
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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)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)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.
Datasheets and documents
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SDS download
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Datasheet download
References (2)
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