TopI-NucEx Kit


SKU: TG1040-1 Categories: ,


Topoisomerase Type I Nuclear Extraction/Assay Kit Product Description

This Kit is designed to allow a researcher to prepare a nuclear extract enriched for type I topoisomerase (topo I) activity and intact 100 kDa polypeptide.  The appropriate assay buffer specific for topo I is included.  This kit is suitable for most animal cell lines growing in vitro but is optimized for human and rodent species. This Kit also allows the investigator to quantify topo I activity in any given cell line and provides the end user a source of enzyme for preliminary drug screening or HTS.


Kit Contents

1.  10x TEMP Buffer.  (10 mM Tris-HCl (pH 7.5), 1 mM EDTA, 4 mM MgCl2, 0.5 mM PMSF).  The kit includes 10 mL (in a 15 mL conical tube).

2.  10x TEP Buffer.  (10 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.5 mM PMSF).  A microfuge tube with 1 mL is provided.

3.  1 M NaCl for extraction (0.5 mL in microfuge tube).

4.  pHOT1 DNA (25 ug total), supercoiled substrate provided at a concentration of 0.25 ug/ml.  DNA concentration may vary in some cases; therefore verify DNA concentrations by referring to the tube label.

5.  Marker DNA, relaxed pHOT1; 0.05 ug/ul in 1x gel loading buffer.

6.  10x Topo I Reaction Buffer (10x TGS; 300 ul).

7.  10x gel loading buffer (300 ul).

8.  10x Proteinase K (500 ul).

9.  10% SDS (300 ul).


Shipping and Storage Instructions:

The kit may be shipped at ambient temperature or on ice (dry ice or wet ice) and should be stored at -20° C upon receipt. Avoid frequent freeze/thaw cycles which may cause DNA nicking. The kit may be store for up to 1 month at 4° C.