The entire multiplex protocol, from labeling to imaging, is completed within an hour. StreptaClick®-antibody conjugates can be combined with DAPI, other directly conjugated primary antibodies or conventional secondary antibodies in the same assay. However, StreptaClick® is not compatible with other streptavidin-based techniques.
Each color contains 500 μl of StreptaClick® antibody labeling reagent, capable of labeling 50 μg of biotinylated antibody, sufficient for approximately 50 slides. The kit also includes a biotin block buffer. This buffer is used to block any remaining free StreptaClick® when multiplexing.
|Name||Absorption max||Emission max|
|AZDye™ 488 – Green||494 nm||517 nm|
|Atto™ 542 -Yellow||542 nm||562 nm|
|AZDye™ 594 – Red||590 nm||617 nm|
|AZDye™ 647 – Far red||649 nm||671 nm|
Step-by-Step Multiplex Staining Protocol
Read the comprehensive user manual for detailed instructions.
Should you biotinylate the antibody ‘in house’ using a biotinylation kit, excess free biotin must be removed, potentially via a spin column, before the labeling process.
- Add the desired amount of biotinylated antibody to be labeled to a suitably sized tube.
- Add the StreptaClick® antibody labeling reagent.
- Incubate for 5 minutes
- Mix all the antibody-StreptaClick® conjugates in one tube with biotin block buffer.
- Apply the mix to your sample and incubate.
- Wash, mount, and analyze under a fluorescence microscope.
Storage and Handling
Shipped in room temperature
Store at 4°C, protect from light, do not freeze.
Note: The StreptaClick® antibody labeling reagent contains 0.09% sodium azide.