HeLa CoCl2 treated/Untreated Cell Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-36450

Novus Biologicals, part of Bio-Techne
Catalog #
Availability
Size / Price
Qty
NBP2-36450
Print Quote
Bulk Order
100% Guarantee

Key Product Details

Species

Human

Applications

SDS-PAGE

Product Summary for HeLa CoCl2 treated/Untreated Cell Lysate

Product Specifications

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).

Type

Cell

Tissue

Cervix

Subcellular Fraction

CoCl2 treated/Untreated

Scientific Data Images for HeLa CoCl2 treated/Untreated Cell Lysate

Western Blot: HeLa CoCl2 treated/Untreated Cell Lysate [NBP2-36450]

Western Blot: HeLa CoCl2 treated/Untreated Cell Lysate [NBP2-36450]

Western Blot: HeLa Hypoxic Simulated / Normoxic Tissue Lysate [NBP2-36450] - Western blot analysis of HIF-1 alpha antibody (NB100-134) in HeLa cobalt chloride treated and normoxic lysates (NBP2-36450). Tubulin loading control is shown below.

Kit Contents for HeLa CoCl2 treated/Untreated Cell Lysate

  • 0.1 ml of HeLa Cobalt Chloride treated whole lysate
  • 0.1 ml of HeLa normoxic whole lysate

Formulation, Preparation, and Storage

Preparation Method

HeLa 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 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.

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: HeLa

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.

Additional HeLa Products

Product Documents for HeLa CoCl2 treated/Untreated Cell Lysate

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for HeLa 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.

Privacy and Cookie Policy
Site Map
© Copyright 2024 Bio-Techne. All Rights Reserved.