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Cell, Acid & Tissue Extracts

COS-7 nuclear extract (CoCl2 treated)

Catalog No: 40600 Format: 200 µg $165 Buy Now

for COS-7 nuclear extract (CoCl2 treated)Contents

2 x 100 µg of COS-7 nuclear extract (CoCl2 treated) at 2.5 µg/µl.

for COS-7 nuclear extract (CoCl2 treated)Background

COS-7 nuclear extract (CoCl2 treated) was prepared from subconfluent undifferentiated cell cultures of the Simian kidney fibroblast COS-7 cell line. The COS-7 cell line was derived from immortalizing the African green monkey CV-1 cell line by transformation with an origin-defective mutant of the SV40 viral genome that is replication-deficient but produces wild-type large T antigen. The large T antigen supports replication of and lytic growth of the SV40 virus as well as replication of SV40 promoter-containing plasmids. In addition to supporting propagation of the Simian SV40 virus, the COS-7 cell line is most commonly used in research related to replication and in transfection studies for the production of recombinant proteins.

Treatment of COS-7 cells with CoCl2 induces hypoxia. Hypoxia is a significant contributor to various developmental and pathophysiological processes. These include cell differentiation, wound healing, angiogenesis and various human diseases, including ischemia, cancer, heart disease pulmonary disease, and inflammation. Hypoxic conditions induce an increase in HIF-1, a transcription factor involved in mammalian oxygen homeostasis. This leads to upregulation of transcription of genes that promote cell survival in low-oxygen conditions, mainly the PI3K/Akt pathway that is important for cell survival. In addition, hypoxia induces proliferation and differentiation of various differentiating cell types, particularly those related to the neural system and vasculogenesis.

for COS-7 nuclear extract (CoCl2 treated)Application Notes

COS-7 nuclear extract (CoCl2 treated) is specifically recommended for 1) analysis of replication, 2) for transfection studies and 3) for studies related to hypoxia.

for COS-7 nuclear extract (CoCl2 treated)Extract Origin

Monkey Kidney, SV40 transformed

for COS-7 nuclear extract (CoCl2 treated)Extract Composition

COS-7 nuclear extract was collected in Lysis Buffer after a 16-hour incubation with CoCl2 (0.15 mM). The Lysis Buffer consists of 20 mM Hepes pH 7.5, 400 mM NaCl, 20% glycerol, 0.1 mM EDTA, 10 mM NaF, 10 µM Na2MoO4, 1 mM NaVO3, 10 mM PNPP, 10 mM β-glycerophosphate, 1 mM DTT and protease inhibitors. The protein content has been determined by a Bradford-based assay.

for COS-7 nuclear extract (CoCl2 treated)Quality Control

Each lot has been tested for HIF activation by using TransAM® HIF Kits. The signal intensity for HIF activation in each lot is compared to the signal intensity obtained with extracts from unstimulated COS-7 cells (see figure). After the signals are blanked, the ratio of the signals from stimulated cells over unstimulated cells must be above 4. This ratio may vary depending on the basal level of HIF activation in a given cell type.

 
Figure 1: Measurement of HIF-1α.

Figure 1: Measurement of HIF-1α.
Nuclear extracts were isolated from COS-7 cells using the Nuclear Extract Kit. The cells were either treated for 20 hours with 0.15 mM CoCl2 or were untreated. Increasing amounts of nuclear extract are assayed for HIF-1α activation using the TransAM HIF-1 Kit.

for COS-7 nuclear extract (CoCl2 treated)Storage

To ensure stability, extracts should be stored at -80°C.

We recommend aliquoting the extracts into single-use fractions and then storing them at -80°C. This eliminates repeated freeze/thaw cycles.

for COS-7 nuclear extract (CoCl2 treated)Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

 

Technical Data Sheet

COS-7 nuclear extract (CoCl2 treated)
 

Data Thumbnails

Figure 1: Measurement of HIF-1α.

Assay of HIF-1α activation using the TransAM HIF-1 Kit
(Click image to enlarge and see details.)