In some cases, I believe the Miltenyi beads can have a detrimental effect on the cells. The bioCapture beads, however, I comepletely inert.

Thanks George for your comments.

In magnetic cell cell sorting using cell isolation kit[+ ve selection] of miltenyi microbeads

they interfere with funtional activity of selected cells'

thank you

dr.deepak.y

Flushing can damage the cells. I had this problem some time ago when trying to capture intact cells from a urine sample. The bioCapture beads work without having to detach them - they can go through the rPCR process right along with the cells and do not affect the assay.

This depends on the downstream application for the cells. If you are selecting them for PCR then the effects of the bound beads should be mininmal. However, the beads do have an effect on cell phenotype that differs depending on cell type. Certainly the bead is bound to the cell by a ligand- receptor interaction. This can, but not neccesarily, trigger signaling through that receptor effecting the phenotype of your isolated cells. In addition some cells, particularly macrophages and dendritic lineage cells, have increased potential to engulf the beads leading to activation of the cells.

You are correct. The beads can effect phenotype assays, but they generally do not have any impact on running PCR based assays.

If I understand the question correctly, then we are talking about functionality of the cells after isolation. As Michael Vetter correctly mentioned it depends on different questions that has to been answered before being possible to answer the question in a satisfying way.

1. What kind of cell will be isolated

2. From which sample material

3. What kind of pre-processing has been done (e.g. Ficoll gradient etc.)

4. Which kind of separation system will be used (Miltneyi, Dynal, Stem Cell Technology, pluriSelect etc.)?

5. Which Ab clone is used with the isolation system

6. What kind of downstream experiments are planed (e.g. activation, phagocytosis, long time culture, differentiation etc.

I you can give me a little more information I can provide you with some answers!?

In case these are sensitive feel free to contact me via e.mail.

sorry spelling mistake:

If you can give me a little more information I can provide you with some answers!?

Did someone try to use MACS sorted cells (positive selection) to make protein IP or ChIP using magnetic beads (Dynabeads)? In my case, I have observed a huge unspecific binding when using MACS sorted cells for ChIP, for example I bring down about 5ug of DNA after ChIP experiments (usually ChIP experiments give few ng of DNA). Is it possible that the Miltenyi microbeads can interfere with immunoprecipitation process when using additional magnetic beads for the IP or ChIP experiments? Thanks by advance for your help.

Does anyone have the experience of isolating some human PBMC subsets (CD4+ T cells, NK, Macrophage) by positive selection using microbeads and then using them for in vivo experiments (injecting into NSG mouse)? Do microbeads affect their function? Thank you.