HepG2 CoCl2 treated/Untreated Cell Lysate

Name: HepG2 CoCl2 treated/Untreated Cell Lysate
Brand: Novus Biologicals
SKU: NBP2-36451

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-36451

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Product Specifications
Scientific Data
Kit Contents
Preparation & Storage
Background
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Specific Notices

Key Product Details

Species

Human

Applications

SDS-PAGE

Product Specifications

Application Notes

HepG2 Whole cell lysate is provided as a Western blot positive control for Hif-2a 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). Boiling before loading is not necessary.

Type

Cell

Tissue

Liver

Subcellular Fraction

CoCl2 treated/Untreated

Scientific Data Images for HepG2 CoCl2 treated/Untreated Cell Lysate

Western Blot: HepG2 CoCl2 treated/Untreated Cell Lysate [NBP2-36451]

Western Blot: HepG2 Hypoxic Simulated / Normoxic Tissue Lysate [NBP2-36451] - Western blot analysis of HIF-2 alpha antibody (NB100-122) in HepG2 cobalt chloride treated and normoxic lysates (NBP2-36451). Tubulin loading control is shown below.

Kit Contents for HepG2 CoCl2 treated/Untreated Cell Lysate

0.1 ml Cobalt Chloride treated HepG2 whole cell lysate
0.1 ml normoxic HepG2 whole cell lysate

Formulation, Preparation, and Storage

Preparation Method

HepG2 cells are cultured using standard media condition until semi-confluent (70-80%). The cells are then treated with or without 100 uM CoCl2 for 4 hours. The cells are then washed in PBS and lysed into 1X Laemmli sample buffer with betaME. The lysate is sonicated and tested by Western blot for reactivity to Hif-2a using a polyclonal anti-Hif-2a antibody (NB100-122). Tubulin reactivity of each lysate is shown as a loading control.

Formulation

The protein lysate is prepared in 1x Laemmli sample buffer with BME. Boiling before loading is not necessary.

Concentration

1.0 mg/ml

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: HepG2

Cobalt Chloride (CoCl2) treated cells are often used as a positive control for HIF-1 and HIF-2 alpha analysis in hypoxia signaling experiments. HIFs induction can be conveniently achieved in vitro through CoCl2 treatment of cultured cells. CoCl2 inhibits prolyl-hydroxylases (PHD enzymes), the oxygen sensors, through replacement of Fe with Co making these enzymes unable to mark HIF alpha for degradation. In other words, CoCl2 allosterically blocks PHDs through its metal ion binding domain, hence, abolishing the VHL-HIF interaction which ultimately leads to the accumulation of HIFs in CoCl2 treated cells. CoCl2 has been regarded as one of the most reliable HIF-1 alpha inducer or hypoxia response mimicker, and this chemically induced hypoxia model is widely used in hypoxia-signaling research. Many researcher use Novus 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 HeLa cells treated with and without CoCl2.

Alternate Names

CoCl2

Additional HepG2 Products

Product Documents for HepG2 CoCl2 treated/Untreated Cell Lysate

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Product Specific Notices for HepG2 CoCl2 treated/Untreated Cell Lysate

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.

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