SUMOstar affinity purification with nanoCLAMP SMT3-A1(Resin), batch mode from E. coli lysate


Overview
Abstract
SUMOstar affinity purification with nanoCLAMP SMT3-A1 by batch mode from over-expressing E. coli lysate.
Type
  • Affinity Chromatography
Specimen
E. coli BL21(DE3)
Target
SUMOstar
Scores
Outcome
Successful
Signal
Acceptable
Cross Reactivity
Acceptable
Background
Acceptable
Dynamic Range
Acceptable
Results
Figure

Image 

SUMOstar affinity purification

Legend 

SDS-PAGE analysis of one step purification of SUMOstar from E. coli BL21DE3 whole cell lysate using SMT3-A1(Resin) affinity resin. E. coli overexpressing SUMOstar were lysed by sonication, cleared by centrifugation, and incubated with 1.8 ml (packed vol) SMT3-A1(Resin) for 1 h at 4°C.  Beads were washed with PBS and eluted with polyol elution buffer. The eluate was buffer exchanged into Mops buffered saline, pH 7.4. Lysates and eluates were analyzed on 12% SDS-PAGE in reducing SDS sample buffer.

Lane 1: Lysate (16 ul) containing overexpressed SUMOstar.

Lane 2: Flow-through (16 ul) post-incubation on 10 ul SMT3-A1(Resin)

Lane 3: 10 ug of buffer-exchanged eluate from SMT3-A1(Resin), from a total of 6.76 mg total elution. 

Note 

This protocol was scaled up to accommodate lysate from 100 ml E. coli expression culture.
Methods
Protocol
Affinity purification from E.coli lysates using nanoCLAMP(Resin)
Product Lot(s) Used
Lot Function Academic Price Commercial Price Order Equivalent Product
nC-SMT3-A1(Cys)10nmol-Prototype Capture $275 $525
Conjugate(s) Used
Conjugate
nanoCLAMP SMT3-A1(Resin)