Calmodulin-Sepharose Beads (ab286869)
Overview
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Product name
Calmodulin-Sepharose Beads -
General notes
Calmodulin-Sepharose beads (ab286869, 7934) allow for specific enrichment of known or unknown calmodulin-binding proteins from biological samples. In addition, Calmodulin-Sepharose beads provide a convenient tool for affinity purification of calmodulin binding peptide (CBP) tagged recombinant proteins. Elution of CBP tagged proteins could be achieved by simply adding chelation ligand / removing Ca2+.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Constituent: 20% Ethanol -
Concentration information loading...
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab286869 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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AP |
Use at an assay dependent concentration.
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Notes |
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AP
Use at an assay dependent concentration. |
Images
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293F cells were lysed with Lysis Buffer (50 mM Tris pH 7.4, 150 mM NaCl, 0.1% Triton X-100, 2 mM CaCl2 and protease inhibitors). Clear lysate was collected after centrifugation at 16,000 g for 30 min at 4˚C. Calmodulin-Sepharose beads (CaM) and Plain beads (Plain) were washed with dH2O, and equilibrated with Wash Buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 2 mM CaCl2). Approximately 25 mg clarified cell lysate (5.8 mg/ml) were incubated with 1 ml of Plain beads or CaM beads at 4˚C
for 2 hours. After incubation, beads were extensively washed with Lysis Buffer, Wash Buffer, and finally with 500 mM NaCl in Wash Buffer. Beads were first eluted with 5 mM EGTA. Equal proportions (20%) of concentrated EDTA eluent were resolved on SDS-PAGE gel (EGTA, 2, 3). The proteins remaining bound to beads were eluted by boiling in 1x SDS loading buffer, and equal proportion of samples (5%) were used for SDS-PAGE (Boiling, 9, 10). Elution of proteins by EGTA and boiling is suggestive of Ca2+-dependent interaction (2, 3) and Ca2+-independent interaction (9, 10) respectively. This clearly indicates that large
populations of proteins are captured by CaM beads (3 & 10), relative to the plain beads (2 & 9). Proteins eluted from CaM beads clearly display a distinct pattern from those in cell lysate (lane 5, 6, 7), supporting the enrichment of specific populations of proteins from lysates.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab286869 has not yet been referenced specifically in any publications.