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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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
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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
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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
