Further information:
QIAexpress
Detection Systems
For convenient molecular weight determination of 6xHis-tagged proteins on western blots
| Sizes of marker proteins: | 15, 30, 50, 75, and 100 kDa |
| Epitope: | N-terminal RGS(H)6 |
| Applications: | Size determination and positive control for western blotting |
| Detection using: | Anti¡¤His Antibodies, Ni-NTA Conjugates |
| Form: | Lyophilized; sufficient for 50¨C100 lanes on western blots |
The 6xHis Protein Ladder serves as a molecular weight
standard as well as a positive control for western blotting of 6xHis-tagged
proteins (see figure "6xHis-Tagged DNA Polymerase Detected
by Tetra¡¤His Antibody in Huh7 Cells"). Five 6xHis-tagged proteins with
apparent molecular weights of 15, 30, 50, 75, and 100 kDa are provided premixed
to yield bands of similar intensity on western blots. They are detected in
parallel with the 6xHis-tagged proteins of interest using QIAexpress
Anti¡¤His Antibodies and HRP Conjugates or Ni-NTA Conjugates. By eliminating the
need to cut off and separately stain gel lanes containing
nontagged markers, the 6xHis Protein Ladder saves time and simplifies western
blotting of 6xHis-tagged proteins.
6xHis-Tagged DNA Polymerase Detected by Tetra¡¤His Antibody in Huh7 Cells
|
The indicated amounts of purified 6xHistagged DNA polymerase mixed with crude extract from Huh7 cells (5 µg total protein per lane) were applied to a 10% SDS-polyacrylamide gel. After electrophoresis and western transfer, 6xHis-tagged proteins were detected with Tetra¡¤His Antibody followed by chemiluminescent detection with AP-conjugated rabbit antimouse IgG and CDP-Star detection reagent. 6xHPL: 6xHis Protein Ladder. |
Sizes and amounts of proteins in the 6xHis Protein Ladder
| Molecular weight |
Amount
per lane (ng)* |
| 100 | 75 |
| 75 | 60 |
| 50 | 50 |
| 30 | 50 |
| 15 | 75 |
| * |
When run on a mini gel as in the QIAexpress Detection and Assay Handbook (2.5 µl per lane). |
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