EasySep Dir Hu CTC Enrich Kit

From STEMCELL Technologies, this kit enriches circulating tumor cells (CTCs) directly from human whole blood by immunomagnetic negative selection. Unwanted hematopoietic cells and platelets are targeted for removal with antibodies recognizing CD2, CD14, CD16, CD19, CD45, CD61, CD66b, and Glycophorin A surface markers, then labeled with EasySep™ Direct RapidSpheres™ and separated using an EasySep™ magnet. Desired cells are simply collected into a new tube and remain untouched, immediately available for downstream applications such as flow cytometry, cell culture, or DNA/RNA extraction.

Designed for processing 100 mL of whole blood, the workflow delivers > 99.9% red blood cell depletion without the need for density gradient centrifugation, sedimentation, or lysis, and up to 3 log depletion of normal hematopoietic cells. The protocol is fast, easy-to-use, and column-free. The kit is compatible for use with most samples, including breast cancer, small cell lung cancer (SCLC) and pancreatic carcinoma samples, and does NOT contain anti-CD36 or anti-CD56.

Typical data from whole blood from healthy donors spiked with approximately 1% CAMA cells (epithelial tumor cell line) show a CTC (epithelial cell+) content of 79 ± 16 % in the non-lysed final enriched fraction (using the silver "Big Easy" EasySep™ Magnet; gated on DRAQ5™ for nucleated cells), with a log depletion of targeted CD45+ cells of 2.8 to 3.2. In an example where CAMA cells were seeded at a starting frequency of 0.98%, the CAMA cell (epithelial cell+) content of the enriched fraction reached 92.02% with a 3.8 log depletion of CD45+ cells. For optimal RBC depletion, blood should be collected using heparin or acid citrate dextrose (ACD); K2EDTA or K3EDTA is not recommended, and unprocessed human whole blood gives the best recovery. Recommended medium is EasySep™ Buffer or PBS containing 2% fetal bovine serum (FBS) and 1 mM EDTA. Components are stored at 2 - 8°C and should not be frozen.