![Western Blot: HeLa Hypoxic / Normoxic Tissue Lysate [NBP2-36452] - Western blot analysis of HIF-1 alpha antibody (NB100-134) in HeLa hypoxic and normoxic lysates (NBP2-36452). Tubulin loading control is shown below.](http://images.novusbio.com/fullsize/HeLa-Hypoxic---Normoxic-Tissue-Lysate-Western-Blot-NBP2-36452-img0002.jpg "Western Blot: HeLa Hypoxic / Normoxic Tissue Lysate [NBP2-36452] - Western blot analysis of HIF-1 alpha antibody (NB100-134) in HeLa hypoxic and normoxic lysates (NBP2-36452). Tubulin loading control is shown below.")
| Reactivity | HuSpecies Glossary |
| Applications | PAGE |
| Concentration | 1.0 mg/ml |
| Preparation Method |
HeLa cells are cultured using standard media condition until semi-confluent (70-80%). The cells are grown under true hypoxic conditions where the oxygen level in the incubator is reduced to 2% for 4 hours to accumulate Hif-1a within the cells. The cells are then washed in PBS and lysed into 1X Laemmli sample buffer with BME. The lysate is sonicated and tested by Western blot for reactivity to Hif-1a using a polyclonal anti-Hif-1a antibody (NB100-134). Tubulin reactivity of each lysate is shown as a loading control. |
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| Application Notes | HeLa Whole cell lysate is provided as a Western blot positive control for Hif-1 research. The lysates are provided as a set of treated and untreated samples. Use 10 ul per lane for a standard mini-gel blot (approx. 1 mg/ml). |
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| Storage | Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles. |
| Buffer | The protein lysate is prepared in 1x Laemmli sample buffer with BME. Boiling before loading is not necessary. |
| Concentration | 1.0 mg/ml |
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