Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were treated with Cobalt Chloride / CoCl2 (Lane 1) or were left untreated (Lane 2) before ...read more
Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were left untreated (C) or were treated with Cobalt Chloride / CoCl2 (H) before preparation ...read more
Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were treated with Cobalt Chloride / CoCl2 (Lane 1) or were left untreated (Lane 2) before ...read more
NB800-PC26 packaging includes 0.2 mg untreated and 0.2 mg treated of COS-7 nuclear extract. NB800-PC26 COS-7 nuclear extract was collected in Lysis Buffer after a 16-hour incubation with CoCl2 (0.15 mM).
Specificity
NB800-PC26 contains hypoxia induced and uninduced COS-7 nuclear extract lysate
Applications/Dilutions
Dilutions
Western Blot
Application Notes
Extracts have been quality control tested by Western blot. Before use mix 1:1 with 2X Sample Buffer and heat to 90C for 5 minutes before running on gel.
Publications
Read Publications using NB800-PC26 in the following applications:
20 mM Hepes (pH 7.5), 400 mM NaCl, 0.1 mM EDTA, 10 mM NaF, 10 uM Na2MoO4, 1 mM NaVO3, 10 mM PNPP, 10 mM beta-glycerophosphate, 1 mM DTT and protease inhibitors.
Preservative
20% glycerol
Concentration
2.5 mg/ml
Lysate Details for Array
Type
Cell
Tissue
Digestive/Waste
Subcellular Fraction
Nuclear Hypoxic Induced
Notes
NB800-PC26 packaging includes 0.2 mg untreated and 0.2 mg treated of COS-7 nuclear extract. NB800-PC26 COS-7 nuclear extract was collected in Lysis Buffer after a 16-hour incubation with CoCl2 (0.15 mM).
Background
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.
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 COS-7 Lysate (NB800-PC26)(8)
We have publications tested in 1 confirmed species: Human.
Tam, KKY. The role of hypoxia inducible factors in regulating ovarian function. Thesis (PhD) University of Adelaide, School of Paediatrics and Reproductive Health, 2010. 2010-01-01 (WB)
FAQs for COS-7 Lysate (NB800-PC26). (Showing 1 - 1 of 1 FAQs).
I am interested in the above product with catalog number NB800-PC26 for western blot. Can I get more information about how the lysates are being produced before proceed with the decision of purchasing? It will be good if you have any protocols and reagents used particularly in western blot application with this product.
NB800-PC26 packaging includes 0.2 mg untreated and 0.2 mg treated of COS-7 nuclear extract. NB800-PC26 COS-7 nuclear extract was collected in Lysis Buffer after a 16-hour incubation with CoCl2 (0.15 mM). These cells come in 20 mM Hepes (pH 7.5), 400 mM NaCl, 20% glycerol, 0.1 mM EDTA, 10 mM NaF, 10 uM Na2MoO4, 1 mM NaVO3, 10 mM PNPP, 10 mM beta-glycerophosphate, 1 mM DTT and protease inhibitors as a buffer.
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![Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were treated with Cobalt Chloride / CoCl2 (Lane 1) or were left untreated (Lane 2) before preparation of lysates (catalog# NB800-PC26). The blot was developed using HIF-1 alpha antibody (catalog# NB100-449).](http://images.novusbio.com/fullsize/COS-7-Nuclear-Hypoxic-Induced-Cell-Lysate-Western-Blot-NB800-PC26-img0003.jpg "Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were treated with Cobalt Chloride / CoCl2 (Lane 1) or were left untreated (Lane 2) before preparation of lysates (catalog# NB800-PC26). The blot was developed using HIF-1 alpha antibody (catalog# NB100-449).")
![Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were left untreated (C) or were treated with Cobalt Chloride / CoCl2 (H) before preparation of lysates (catalog# NB800-PC26). The blot was developed using HIF-1 alpha antibody (clone H1alpha67; catalog# NB100-105).](http://images.novusbio.com/fullsize/COS-7-Nuclear-Hypoxic-Induced-Cell-Lysate-Western-Blot-NB800-PC26-img0001.jpg "Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were left untreated (C) or were treated with Cobalt Chloride / CoCl2 (H) before preparation of lysates (catalog# NB800-PC26). The blot was developed using HIF-1 alpha antibody (clone H1alpha67; catalog# NB100-105).")
![Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were treated with Cobalt Chloride / CoCl2 (Lane 1) or were left untreated (Lane 2) before preparation of lysates (catalog# NB800-PC26). The blot was developed using HIF-1 alpha antibody (catalog# NB100-479).](http://images.novusbio.com/fullsize/COS-7-Nuclear-Hypoxic-Induced-Cell-Lysate-Western-Blot-NB800-PC26-img0002.jpg "Western Blot: COS-7 Nuclear Hypoxic Induced Cell Lysate [NB800-PC26] - WB analysis of 50ug nuclear lysate of COS7 cells which were treated with Cobalt Chloride / CoCl2 (Lane 1) or were left untreated (Lane 2) before preparation of lysates (catalog# NB800-PC26). The blot was developed using HIF-1 alpha antibody (catalog# NB100-479).")