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[독일 IBA] Detection of Strep-tag® fusion proteins
엘바이오시스템
Western blot
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)
ELISA
For protein detection in ELISA use
StrepMAB-Classic, unconjugated
Strep-Tactin® HRP conjugate
Immunocytochemistry/Immunohistochemistry
Visualize your Strep-tag® protein with the following dyes:
Strep-Tactin® , StrepMAB-Classic and StrepMAB-Immo are available with single fluorescent conjugates at different wavelengths (488 nm, 546 nm, 645 nm):
Strep-tag® fluorescent Chromeo 488 conjugates of Strep-Tactin®, StrepMAB Classic and StrepMAB Immo (50 µg each)
Strep-tag® fluorescent Chromeo 546 conjugates of Strep-Tactin®, StrepMAB Classic and StrepMAB Immo (50 µg each)
Strep-tag® fluorescent Oyster 645 conjugates of Strep-Tactin®
(50 µg)
FACS
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.
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