Seahorse XF24, Руководство по обучению

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Basic Procedure
12
Preparation of Assay Media for Use in XF Assays
This basic procedure details the preparation of assay media for use with
(1) XF Cell Mito Stress Test Kit, and (2) XF Glycolysis Stress Test Kit
Seahorse recommends the use non-buffered assay medium for XF assays to ensure accurate, functional
measurements of metabolic phenotypes in an ambient environment. The optimal assay media for the
XF Cell Mito Stress Test and the XF Glycolysis Stress Test are different. Both can be prepared starting
with the XF Base Medium and adding different substrates as determined by your cell model. Substrate
requirements are cell type specific and may need to be determined empirically. For the XF Cell Mito
Stress Test, assay medium with user-added glucose, sodium pyruvate and glutamine is recommended
(see 1). For the XF Glycolysis Stress Test, assay medium with user-added glutamine is recommended
(see 2).
1. XF Cell Mito Stress Test Assay Medium
For one tissue culture plate: if rinsing manually, 100 mL of assay medium is sufficient; if rinsing using
the XF Prep Station 200 mL of assay medium is sufficient.
Materials
Source Cat #
XF Base Medium Seahorse Bioscience 102353-100
Glucose Sigma G7528
Sodium Pyruvate (powder) Sigma P5280
Sodium Pyruvate (liquid, 100 mM) Sigma S8636
L-Glutamine (200 mM) Life Technologies 25030-081
0.2
µ
M Sterile Filter
pH Meter
1 N NaOH
Method
1. Warm 100 mL XF Base Medium to 37°C.
2. Add glucose for the desired final concentration using the following table.