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In this application protocol, we describe an easy and fast method for the dissociation of skeletal muscle tissue from mouse and a subsequent isolation method that generates a highly pure population of satellite cells and minimizes bias in downstream applications caused by contamination with non-target cells.
(Optional) ART. 1000 REACH™ pipet tips (Molecular BioProducts, Inc.) for removal of dissociated material from the closed C Tubes
|Column||Max. number of labeled cells||Max. number of total cells||Separator|
|LS||2×107||4×107||MidiMACS™, QuadroMACS™, |
VarioMACS, SuperMACS II
▲ Note: Column adapters are required to insert certain columns into the VarioMACS™ or SuperMACS™ II Separators. For details refer to the respective MACS Separator data sheet.
Use the Skeletal Muscle Dissociation Kit to generate a single-cell suspension by combining mechanical dissociation with the gentleMACS™ Dissociator and enzymatic degradation of the extracellular matrix that maintains the structural integrity of the tissue. Follow the protocol of the kit data sheet.
Download data sheet
Isolate functional satellite cells by depleting non-target cells with the Satellite Cell Isolation Kit. The kit contains a cocktail of monoclonal antibodies conjugated with MACS® MicroBeads the magnetically label non-target cells. These are retained within the MACS Column, while the unlabeled satellite cells pass through during elution. Follow the protocol of the kit data sheet.
Download the data sheet
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