Invitrogen™ Dynabeads™ CD45

Invitrogen™ Dynabeads™ CD45

Dynabeads™ CD45 are superparamagnetic beads covalently coupled to anti-human CD45 antibody that enable isolation or depletion of CD45⁺ leucocytes directly from whole blood, buffy coat, or MNC suspensions. Dynabeads™ CD45 can also be used to enrich epithelial tumor cells. The CD45 antigen is weakly expressed on myeloid cells. To efficiently deplete or isolate all leucocytes including myeloid cells, use Dynabeads™ CD15 (Cat. No. 11137D) in combination with Dynabeads™ CD45.

About Dynabeads™ CD45, Dynabeads™ CD45 are uniform, superparamagnetic beads (4.5 µm diameter) coated with a primary monoclonal mouse IgG2a antibody specific for a CD45 membrane antigen common to all known isoforms of CD45. CD45 is expressed on all human leucocytes. Because of the bead size, Dynabeads™ CD45 can easily and efficiently isolate or deplete cells from viscous samples such as whole blood and bone marrow in about 30 minutes. Dynabeads™ CD45 is often used for enrichment of circulating non-hematopoietic tumor cells, by depletion of all CD45⁺ leucecytes from a MNC sample. Positively isolated cells can be used for downstream molecular studies; for example, those in which cells are to be lysed while still attached to beads, and nucleic acids or proteins further purified. Note that intact cells will not be released from these beads, and we do not recommend to analyze the bead-bound cells with a flow cytometer.

Magnetic bead–based separation offers easy handling, Dynabeads™ CD45 is added to the sample under continuous mixing to optimize the binding of the Dynabeads™ to the target cells. By placing the sample on a magnet it separates the bead-bound target cells from the rest of the sample in just 1–2 minutes. For depletion, remove the supernatant to a new tube for further studies and discard the bead-bound cells. For positive isolation for molecular studies, remove the supernatant and wash the bead-bound cells 2–3 times in buffer to get optimal purity. The cells can be lysed while still attached to the beads and the supernatant transferred to a new tube for downstream molecular analysis. Starting samples can be whole blood, buffy coat, bone marrow, PBMC, or tissue digests.

Features

• Efficient depletion of human leucocytes.
• Isolation of leucocytes directly from whole blood for molecular applications.
• Enrichment of circulating epithelial tumor cells.

Contents

• 5 ml Dynabeads™ CD45 coated with anti-CD45 monoclonal antibody.