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HeLa Hypoxic / Normoxic Cell Lysate

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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.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications PAGE
Concentration
1.0 mg/ml

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HeLa Hypoxic / Normoxic Cell Lysate Summary

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.

Applications/Dilutions

Dilutions
  • SDS-Page 10 ul / lane
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).
Publications
Read Publication using NBP2-36452.

Packaging, Storage & Formulations

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

Lysate Details for Array

Type
Cell
Tissue
Cervix
Subcellular Fraction
Hypoxic / Normoxic

Kit Components

Components
  1. 0.1 ml Hela hypoxic whole cell lysate
  2. 0.1 ml Hela normoxic whole cell lysate

Background

Hypoxia contributes significantly to the pathophysiology of many Human diseases. Many researcher use antibodies to Hif-1 and Hif-2 in Western blot analysis as markers for Hypoxia. For this reason it is important to have reliable positive controls for your Western blot to ensure that your antibodies and test conditions are optimized. Novus Biologicals is pleased to offer the same positive control cell lysates used to validate antibodies to Hif-1 and Hif-2 to our customers for their research. For Hif-1, we have matched pairs of lysate from true HeLa normoxic and hypoxic lysates.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Publications for HeLa Lysate (NBP2-36452)(1)

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