Detection of Strep-tag® fusion proteins
For direct detection of Strep-tag® fusion proteins in Western blots
- via chromogenic and/or chemiluminescence reaction (no secondary antibody needed!) use
- Strep-Tactin® HRP conjugate (HRP, horseradish peroxidase)
- StrepMAB-Classic HRP conjugate
- via chromogenic reaction
- Strep-Tactin® AP conjugate (AP, alkaline phosphatase)
For protein detection in ELISA use
- StrepMAB-Classic, unconjugated
- Strep-Tactin® HRP conjugate
Visualize your Strep-tag® protein with the following dyes:
Strep-Tactin®XT, StrepMAB-Classic and StrepMAB-Immo are available with single fluorescent conjugates:
- Strep-tag® fluorescent Dy488 conjugates of Strep-Tactin®XT, StrepMAB-Classic and StrepMAB-Immo (50 µg each)
For staining (Twin-)-Strep-tagged proteins in FACS we recommend to use our Strep-Tactin® PE 6-5001-005 or Strep-Tactin® APC 6-5011-005 conjugates, because they lead to a more intensive staining compared with our Oyster and Chromeo dyes and are therefore better suited for a light sensitive application as FACS.
- Wash 5x106 pre-cooled cells with 10 ml of a biotin-free physiological wash buffer in a 15 ml reaction tube to remove potentially interfering ingredients (e.g. biotin).
- Resuspend the cells in 50 μl wash buffer and transfer cells to a reaction vessel suitable for staining, e.g. a 96-well round bottom microtiter plate.
- Add 5 µl Strep-Tactin®PE (or Strep-Tactin®APC) to the cells and mix thoroughly by gentle pipetting.
- Incubate for 20 minutes at 4°C in the dark.
- Wash cells three times (400xg, 2 min) in 200 μl wash buffer.
- Cells are ready for FACS
Note: The staining intensity depends on the concentration of tagged protein and in case of unsatisfying results the concentration of conjugate has to be adjusted.