Frequently Asked Questions
Yes, we offer
- 8-16% for mild-range separations
- 4-20% for wide-range seperations
No, NuSep gels are compatible with most popular mini-gel tanks.
- NN Cassette – 10cm x 10cm – fits XCell Sure Lock tanks (Novac)
- NB Cassette – 10cm x 8.5 cm – fits Mini-Protean tanks (Biorad)
- NG Cassette – 10cm x 8cm – is compatible with most generic tanks
All the gels have a 4% stacking gel.
No, the gels do not contain SDS (SDS is incorporated in the running buffer solution). The gels can be used for native runs.
NuSep gels and sample buffers do not contain reducing agents; they can be added to a sample prior to loading. Beta-mercaptoethanol and DTT are both suitable for use with our gels.Â
*see Running Formulations
Yes, NuSep gels must be air-dried. Use of vacuum drying can result in cracking. NuSep gels can only be dried crack-free with NuSep Gel Drying Solution; all other drying solution can result in cracking. A recommended gel drying protocol can be found on the NuSep website.
Troubleshooting
Cause: Air bubbles in the sample wells, or between the gel and cassette, or at the bottom of the cassette
Solution: Use a transfer pipette to displace the air bubbles from the sample wells
Cause: Poorly soluble or weakly charged particles (such as carbohydrates) in the sample
Solution:
- Centrifuge sample
- Change pH of buffer
- Heat sample in the presence of SDS
Cause: Incorrect gel selection, sample overloading, insufficient cooling buffer
Solution:
- Reduce sample size
- Select a gel which separates in the desired molecular weight range
- For proteins of similar molecular weight, a 2-D separation may be required
- Increase buffer in outer tank
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Cause: Too much salt in the sample
Solution: Reduce salt by dialysis or ultra-filtration
Solution: Enzymatically remove the carbohydrate
Solution: Use gel with large pore size at top, try addition of a non-ionic detergent
Cause: Excessive heating
Solution: Always start will chilled buffer (<15C)
Cause: SDS still present in gel
Solution: Fix in 10% TCA prior to staining, otherwise use 30% methanol in the destaining soution
