I am looking for a molecular weight marker that can be used for BN PAGE, and I am wondering if the product metioned above gives nice sharp bands. I am also open for any other suggestions you might have. The MW range between 100 and 400 kDa is especially important for me.
Dear Tom,
indeed an important issue! The decisive point for selecting a suitable molar mass marker is the nature of your sample proteins, in particular whether those are (predominantly) soluble ones or integral membrane proteins (i.e. protein-lipid-detergent particles). Accordingly, the marker proteins need to reflect those properties as closely as possible. In case of membrane proteins, one can simply use a detergent extract of mitochondria (covering a big MW range by OXPHOS complexes) comprising the same detergent as used in the sample.
The "HMW Native Marker Kit" is suitable for soluble proteins and has been often used, allowing for a good calibration up to about 700 kDa.
We often used the HMW marker for BN-PAGE, and it has a fairly good range, but equally, just using BSA (60kDa), which tends to form dimers and trimers, covers the lower molecular weight range quite well. Also, we used holo-transferrin from Sigma as it has 2 bands, 1 at 400kDa and one at 800kDa, and since it is orange in colour, it is easily visible immediately after the run, so helps to let you know that the gel ran ok. So combining BSA and transferrin might work better for your range.
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