K-562 nuclear extract

Catalog No: 36015 Format: 200 µg $220 Buy

Contents

200 µg of K-562 nuclear extract at 4.3 µg/µl.

Background

K-562 nuclear extract was prepared from a cell culture of the K-562 human myelogenous leukemia cell line. The K-562 cell line was originally derived from immortalized erythroleukemia cells obtained from a patient with a Philadelphia chromosome-positive chronic myeloid leukemia in blast crisis. These pluripotent cells are widely applicable in research due to their ability to spontaneously differentiate into erythrocyte, granulocyte and monocyte progenitors. K-562 cells are also frequently utilized in in vitro natural killer (NK) cell assays as highly sensitive targets since they lack the expression of MHC class I molecules required for protection against NK activity.

Application Notes

K-562 nuclear extract is recommended for use in the studies related to 1) differentiation, 2) immunology, and 3) cancer.

Extract Origin

Human lymphoblast bone marrow; chronic myelogenous leukemia (CML)

Extract Composition

K-562 nuclear extract is supplied in Dilution Buffer (20 mM Hepes (pH 7.9), 100 mM KCl, 1 mM MgCl2, 20% glycerol, 0.5 mM PMSF and 0.5 mM DTT).

Quality Control

Extracts have been quality control tested by Western blot and the Electrophoretic Mobility Shift Assay (EMSA).

 

Figure 1: Monitoring GATA family member activation using the TransAM GATA Family Kit.
GATA-1, -2 and -3 activation were assayed using the TransAM GATA Family Kit. 1:1000 dilutions of each antibody in the kit were tested using 5 µg/well of nuclear extract prepared from an unstimulated cell line: GATA-1 and -2 were tested with K-562 and GATA-3 with Jurkat. Assays were performed in the absence or presence of 20 pmol of competitor oligonucleotide that contains either a wild-type or mutated GATA consensus binding site. Note that the wild-type oligonucleotide reduces GATA binding by over 90%, while incubation with the mutant GATA competitor oligo has a limited effect on GATA-1, -2 and -3 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.