FAQs: SageELF automated 1D protein fractionation

Q: Does the SageELF use PAGE?
A: No, cassettes are pre-cast with agarose and 0.1% SDS.

Q: What is the maximum sample amount that can be fractionated?
A: 350ug. Samples are prepared in a 22 ul volume to which TCEP and loading solution is added. Total volume in 40 ul.

Q: What is the volume of the collected fractions?
A: 28 ul per well.

Q: What is the buffer?
A: Tris TAPS, ph 8.7.

Q: What do you recommend for SDS removal?
A: We recommend the Pierce HiPPR detergent removal spin columns.

Q: What are the fractionation ranges?
A: At present, we offer 3% agarose cassettes (ELP3010) for ranges between 10-300 kDa and 5% agarose cassettes (ELP5010) for ranges between 10-150 kDa

Q: How long can you store a gel cassette?
A: 18 months.

Q: How does the software calibration work?
A: A labelled DNA oligo is added the sample (this is optional). The oligo runs ahead of the protein sample and when it is detected a run-time threshold is set. Software then uses calibration data based on run-time to estimate a fraction value in all 12 wells.

Q: How long is a run?
A: Typically 2-3 hours.

Q: Is the sample reduced and alkylated?
A: Our sample prep requires reducing with TCEP (DTT may also be used). We are evaluating alkylation at this time.

Q: What extraction method works best?
A: Extraction methods that are designed for SDS PAGE analysis are preferred. Extraction methods for IEF using urea/thiourea should be avoided.

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