Hep G2 nuclear extract (IL-6 stimulated, 100 ng/ml)

Catalog No: 36107 Format: 200 µg ¥44,000 Buy

Contents

2 x 100 µg of Hep G2 nuclear extract (IL-6 stimulated, 100 ng/ml) at 2.5 µg/µl.

Background

Hep G2 nuclear extract (IL-6 stimulated, 100 ng/ml) was prepared from cell cultures of the human hepatocellular liver carcinoma Hep G2 cell line. These cells are epithelial in morphology and can be induced to exhibit polarization under appropriate culture conditions. Hep G2 cells are most commonly used for studies of hepatocyte function. Because of their ability to differentiate into polarized epithelium, these cells are routinely utilized to investigate intracellular protein trafficking, particularly in relation to human liver diseases. Hep G2 is the most commonly used cell line in studies pertaining to the regulation of hepatic protein synthesis, particularly the synthesis of acute phase proteins after the onset of a systemic inflammatory response. In addition, these cells are important for studies related to liver metabolism, liver cancers, liver regeneration, liver cytotoxicity, apoptosis and as a model for hepatitis B virus (HBV) viral etiology.

In response to injury, disease or infection, interleukin-6 (IL-6) is a cytokine that is rapidly and transiently expressed as a host defense mechanism. IL-6 has various biological activities, most of which are associated with acute phase immune and inflammatory responses. In the liver, IL-6 is major regulator of acute phase protein synthesis in response to inflammation. Studies have demonstrated that constitutive IL-6 has deleterious effects on the liver and may be involved in the onset of liver disease, pathogenesis and cancer.

Application Notes

Hep G2 nuclear extract (IL-6 stimulated, 100 ng/ml) is specifically recommended for studies associated with inflammation of the liver and related to 1) human liver diseases, cancers and drug therapies, 2) HBV etiology, 3) signaling pathways, and 4) apoptosis.

Extract Origin

Human Liver (hepatocellular carcinoma)

Extract Composition

Hep G2 nuclear extract (IL-6 stimulated, 100 ng/ml) is supplied in lysis buffer consisting 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, 10mM PNPP, 10 mM b-glycerophosphate, 1 mM DTT and protease inhibitors. Cells were cultured in medium supplemented with 100 ng/ml IL-6 (interleukin 6) for 15 minutes at 37°C immediately prior to harvesting.

Quality Control

Each lot has been tested for STAT activation by using TransAM® STAT Family Kits.

 
Figure 1: Monitoring STAT Family member activation using the TransAM STAT Family Kit.

Figure 1: Monitoring STAT Family member activation using the TransAM STAT Family Kit.
STAT1α, STAT3, STAT5A and STAT5B activation were assayed using the TransAM STAT Family Kit. 1:1000 dilutions of each antibody in the kit were tested using 5 µg/well of nuclear extract prepared from a stimulated cell line: STAT1α was tested with U-937 (TPA + IFNγ), STAT3 with Hep G2 (IL-6, 100 ng/ml), and STAT5A and STAT5B with Nb2 (prolactin). Assays were performed in the absence or presence of 20 pmol of competitor oligonucleotide that contains either a wild-type or mutated STAT consensus binding site. Note that the wild-type oligonucleotide reduces STAT binding by over 90%, while incubation with the mutant STAT competitor oligo has a limited effect on STAT 1α, STAT3, STAT5A or STAT5B binding to DNA

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.

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.