Nb2 nuclear extract (prolactin stimulated)

Catalog No: 36106 Format: 200 µg ¥2,860 Buy

Contents

2 x 100 µg of Nb2 nuclear extract (prolactin stimulated) at 2.5 µg/µl.

Background

Nb2 nuclear extract (prolactin stimulated) was prepared from cell cultures of the rat lymphoma Nb2 nuclear cell line. The Nb2 cell line was derived from the lymphoma transplant in the thymus of a noble (Nb) strain male rat following prolonged estrogen treatment. Nb2 lymphoma cells are of pre-T cell origin and express the alpha and beta chains of the T cell receptor. However, they lack expression of class II MHC antigens. These cells are dependent on lactogens for mitogenic activity, which can also be stimulated by cytokines and human growth hormone and its analogs. Nb2 cells are most commonly used in studies of lactogen-induced mitogenesis and as a model for tumor progression of T cell cancers.

Application Notes

Nb2 nuclear extract (prolactin stimulated) is specifically recommended for studies related to lactogen-induced mitogenesis and T cell cancers.

Extract Origin

Rat lymphocytes

Extract Composition

Nb2 nuclear extract derived from rat lymphocytes was collected in Lysis Buffer after a 20 minute stimulation with 25 ng/ml of prolactin at 37°C immediately prior to harvesting. Lysis Buffer consists of 20 mM Hepes pH 7.9, 100 mM KCl, 20% glycerol, 1 mM MgCl2, 0.5 mM DTT and 0.5 mM PMSF. The protein content has been determined by a Bradford-based assay.

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.
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.