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mCherry purification by affinity chromatography with nanoCLAMP mCher-A2(Resin), batch mode from E. coli lysate
| Abstract | mCherry purification by affinity chromatography with nanoCLAMP mCher-A2(Resin) in batch mode from E. coli lysate. E.coli were lysed with BPER (Thermo), cleared by centrifugation, and diluted with PBS to 1.2 mg total protein/ml. The lysate was spiked with recombinant antigen to 0.1 mg/ml in 1.4 ml and incubated with 10 µl (packed vol) mCher-A2(Resin). Beads were washed with PBS and eluted with polyol elution buffer at near neutral pH. |
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| Type |
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| Target |
| Outcome | Successful |
|---|---|
| Signal | Acceptable |
| Cross Reactivity | Acceptable |
| Background | Acceptable |
| Dynamic Range | Acceptable |
| Figure | Image LegendSDS-PAGE analysis of one step purification of mCherry from E.coli BL21DE3 whole cell lysates using mCher-A2(Resin) affinity resin. SDS-PAGE analysis of one-step purification of spiked proteins from E. coli BL21(DE3) whole cell lysates using nanoCLAMP chromatography resin. E. coli were lysed with BPER, cleared by centrifugation, and diluted with PBS to 1.2 mg total protein/ml. The lysate was spiked with recombinant antigen to 0.1 mg/ml in 1.5 ml and incubated with 10 µl (packed vol) nanoCLAMP resin. Beads were washed with PBS and eluted with polyol elution buffer at near neutral pH. The gel was loaded with lysate containing approximately 10 µg total protein (lane 1) or with 8 ul of eluate (from a total of 125 µl eluate) from nanoCLAMP resin, in reducing SDS-sample buffer. Lane 1: mCherry-spiked lysate containing 10 ug total protein. Lane 2: 8 µl of polyol eluant from the affinity resin, from a total of 125 µl total elution.
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