HeLa DNA Damage Whole Cell Lysate Set: UV Treated and Untreated Control (ab157396)
Overview
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Product name
HeLa DNA Damage Whole Cell Lysate Set: UV Treated and Untreated Control -
Species reactivity
Reacts with: Human -
Product overview
UV light is a common source of DNA damage, and can lead to skin cancer and premature aging. Exposure to UV-B and UV-C light leads to the formation of pyrimidine dimers, and to a lesser extent, purine dimers and pyrimidine photophosphates. These dimerized DNA bases are typically removed by the nucleotide excision repair pathway. Failure to repair the damage can induce apoptosis by blocking DNA replication and transcription.
The UV-treated HeLa lysate is designed for use as a western blot positive control when studying UV-induced DNA damage and/or apoptosis. Cells were treated with UV-C light for 1 minute, then cultured for 4 hours before collecting for lysates. Untreated cells were grown under standard cell culture conditions for HeLa cells.
Samples are provided solubilized in an SDS-PAGE loading buffer, supplemented with reducing agent. This sample is ready for SDS-electrophoresis and acts as a positive control in Western blotting applications.
Concentration: HeLa UV-treated lysate, 200 µg at 2.0 mg/mL
HeLa untreated lysate, 200 µg at 2.0 mg/mL -
Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 2 units Control for UV-Treated HeLa Lysate 1 x 200µg UV-Treated HeLa Lysate 1 x 200µg -
Research areas
Images
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Figure 1. Western blot with ab136812 Apoptosis Western Blot CocktailLane 1: MW marker
Lane 2: Untreated HeLa lysate (ab157396)
Lane 3: UV-treated HeLa lysate (ab157396).
All lysates at 20 µg per lane.
Primary antibodies (all lanes): ab136812 Apoptosis Western Blot Cocktail (pro/p17-caspase 3, cleaved-PARP, muscle actin) 1:250 dilution.
Secondary antibodies (all lanes): Goat polyclonal to Mouse IgG - HRP at 1:5000 dilution and Goat polyclonal to Rabbit IgG - HRP at 1:5000 dilution.
Developed using the ECL method.
Predicted band sizes: 89, 42, 32, 17 kDa
Observed band sizes: 89, 42, 32 kDa -
Figure 2. Western blot with ab133981 Anti-Caspase 9 antibodyLane 1: MW marker
Lane 2: Untreated HeLa lysate (ab157396)
Lane 3: UV-treated HeLa lysate (ab157396)
All lysates at 20 µg per lane.
Primary antibody (all lanes): ab133981 Anti-Caspase 9 antibody at 2 µg/mL.
Secondary antibodies (all lanes): Goat polyclonal to Mouse IgG - HRP at 1:5000 dilution.
Developed using the ECL method.
Predicted band size: 35 kDa
Observed band size: 25 kDa
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Figure 3. Western blot with ab131385 Apoptosis and DNA Damage Western Blot CocktailLanes 1, 4: MW marker
Lane 2: Untreated HeLa lysate (ab157396)
Lane 3: UV-treated HeLa lysate (ab157396).
All lysates at 20 µg per lane.
Primary antibodies:
Lanes 1-3: ab131385 Apoptosis and DNA Damage Western Blot Cocktail (cleaved PARP, GAPDH, H2A.X pSer139) 1:250 dilution
Lanes 4-6: H2A.X pSer139 antibody.
Secondary antibodies (all lanes): Goat polyclonal to Mouse IgG - HRP at 1:5000 dilution.
Developed using the ECL method.
Predicted band sizes: 15, 36, 89 kDa
Other bands observed (from H2A.X pSer139 antibody, identities unknown): 25, 30 kDa