Viromer® RED

Protocol tips

Upstream tips	Protocol tips	Downstream tips
For adherent cells, you can target up to 80% confluency at the time of analysis	Always add Buffer RED to Viromer® RED, not vice versa!

Upstream tips
For adherent cells, you can target up to 80% confluency at the time of analysis
Protocol tips
Always add Buffer RED to Viromer® RED, not vice versa!

Publication protocol

Keratinocyte cell culture, transfection and sorting
Primary KCs were harvested from the backs of adult C3H mice at 54 ± 2 days and KC cell lines arising from these cultures were maintained in Morris II medium [34]. Cell lines were periodically tested for Mycoplasma. No other authentication was performed. For transient transfection in HEK293 (ATCC, cat no.SD-3515,) and in the KC cell line, cells were transfected using VIROMER RED (Lipocalyx GmbH, Germany). For ROCK1 knockdown in KCs, cells were transfected with pooled shRNA reagent (All shRNAs were obtained from the BioMedical Genomics Center at The University of Minnesota). The negative control vector was the pLKO.1 vector backbones that has no hairpin insert. Primary KCs were stained with antibodies to CD34 (rat anti-mouse CD34-FITC, BD Pharmingen); rat anti-mouse CD49f-PE [(alpha-6 integrin), BD Pharmingen] and stem cell fraction (CD49f+CD34+) and stem cell depletion fraction (CD49f +CD34−) were sorted using Fluorescence Activated Cell Sorting (FACS) as previously described [28].

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Papers

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Manufacturer protocol

Download the product protocol from Lipocalyx GmbH for Viromer® RED below.

Download manufacturer protocol

Videos

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