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Is it possible to use frozen primary CD34+ hematopoietic stem cells for Nucleofection®?

Yes. For cryopreserved PBMCs or enriched CD34 populations, we recommend culturing the thawed cells 1-2 hours in culture medium before Nucleofection®. Any further enrichment procedure after thawing is not recommended. Viability of frozen nucleofected...
Missing: stable linesGuideline

I thawed and plated my Clonetics™ embryonic rat or mouse neuronal cells and there are a lot of dead cells. What happened?

Cell death will usually be observed during the first few days of growth, resulting in cell debris in the culture – this is normal. Cell cultivation should be continued and surviving cells should start to develop. By day 4, neurite networks...
Missing: stable linesGuideline

What is the difference between the different cell sources (bone marrow, cord blood, or fetal liver) for CD34+ cells?

Bone Marrow: provides a hematopoietic microenvironment and the highest number of different donors.Cord Blood: more naive cells useful in transplant and stem cell research.Fetal Liver: best for developmental studies and they are highly proliferative...
Missing: stable linesGuideline

What are useful methods to enrich/purify specific hematopoetic cell populations before Nucleofection™?

Specific blood cell populations can be enriched/purified for Nucleofection™ by MACS™ selection Kits (Miltenyi Biotec GmbH), RosetteSep™ Kits (StemCell Technologies Inc.) or Dynal™ beads (Invitrogen Corporation).
Missing: stable linesGuideline

How are Lonza's CD34+ cells isolated?

Lonza's CD34+ cells are isolated first by obtaining mononuclear cells from bone marrow or cord blood via density gradient centrifugation. The CD34+ cells are then isolated form the mononuclear cell population using positive immunomagnetic selection.
Missing: stable linesGuideline

What is the purity of Lonza's CD34+ cells?

Cell purity (as determined via FACS analysis for CD34 expression) is guaranteed to be 90% or greater for all Lonza CD34+ cell products. For some Lonza CD34+ cell products, a "budget" version of the product is available. The "budget" versions...
Missing: stable linesGuideline

How to maximize cell viability in a Nucleofection® experiment?

Pay careful attention to centrifugation speeds prior to Nucleofection®; be sure not to centrifuge the cells at higher than 90xg. Any trypsinization should use the minimal amount of reagent and time necessary in order to minimize stress to the cells...
Missing: stable linesGuideline

Can I optimize the transfection efficiency or cell viability after the 4D-Nucleofector® Cell Line Optimization any further?

We offer a program fine tuning matrix for Optimization of Nucleofection® Conditions - A Short Guideline (lonza.com)Based on your current most satisfactory pulse(s) it provides guidelines which further programs to test to either increase cell...
Missing: stable linesGuideline

How can cells be cured of mycoplasma contamination?

Where contamination has occurred, and the sample absolutely cannot be discarded, the MycoZap™ Mycoplasma Elimination Reagent has been optimized to eliminate mycoplasma with minimal toxic effects on the cells. The MycoZap™ Reagent eliminates...
Missing: stable linesGuideline

How can cells be cured of mycoplasma contamination?

Where contamination has occurred, and the sample absolutely cannot be discarded, the MycoZap™ Mycoplasma Elimination Reagent has been optimized to eliminate mycoplasma with minimal toxic effects on the cells.The MycoZap™ Reagent eliminates...
Missing: stable linesGuideline
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