StreptaClick® - Multiplex Immunofluorescence
Multiplex Immunofluorescence made easy
Manual multiplex immunostainings can be time-consuming, but the StreptaClick® technology is here to help. StreptaClick® utilizes monovalent streptavidin’s extraordinary capabilities to rapidly and firmly bind biotinylated antibodies in solution – without causing antibody aggregation. Simply pre-mix biotinylated antibodies with StreptaClick®-HRP or StreptaClick®-color for fluorophores, and obtain directly conjugated antibodies in minutes without wash steps. Multiple StreptaClick®-labeled antibodies can then be used for multiplex IHC.
StreptaClick® – HRP Workflow
How it works
Biotinylated antibodies pre-mixed with StreptaClick® operate similarly to directly conjugated antibodies, thereby eliminating the need for secondary antibodies. The StreptaClick®-labeling reaction is extremely efficient, permitting the labeling of any antibody volume, even down to microliters. This is particularly beneficial when setting up new experiments.
Quick Results, Less Time
StreptaClick®-color enables a multiplex IHC protocol—from labeling to imaging—to be completed within one hour. Each fluorochrome is attached to its biotinylated antibody with a one-step, 5-minute StreptaClick® reaction. All StreptaClick®-labeled antibodies can then be combined and added to the slide simultaneously.
With StreptaClick®-HRP you can complete a 7-color Tyramide Signal Amplification (TSA) multiplex IHC within a single day (DAPI + 6 biotinylated antibodies). Each biotinylated antibody is attached with HRP by the fast StreptaClick® reaction, and the kit includes an HRP block buffer that efficiently quenches HRP activity between staining cycles. This avoids the slow, inconvenient and harsh boiling step to strip off antibodies between staining cycles.
Seven-Color TSA method for frozen sections from mice, humans, rats, and other species.
The StreptaClick®-HRP solution is a must for any manual TSA staining. The kit comes with a quenching buffer that eliminates the need for boiling, which not only accelerates each color cycle but also makes the kit highly beneficial for both FFPE and frozen sections. The quenching buffer effectively silences the HRP signal from the previous round while preserving the morphology of the frozen tissue. Hence, the StreptaClick®-HRP kit provides a rapid method for multiplex TSA IHC on both FFPE and frozen sections, with antibodies from any host, and on tissues from mice, humans, rats, and other species.
All our fluorochromes have been carefully selected for their outstanding brightness and signal intensity to enhance image quality and scanning speed. For TSA, we offer an optional selection of eight optimized tyramide dyes that feature remarkable signal intensity, delivering a signal 10-50x brighter than direct conjugates. StreptaClick®-Color comes with green, yellow, red, and far red fluorochromes (AZDye™ 488, Atto™ 542, AZDye™ 594, and AZDye™ 647). AZDyes™ have the same chemical structure as Alexa Fluor® dyes, while Atto 542 is an exceptionally bright dye with spectral properties similar to Cy®3.
The fluorochromes we provide are carefully chosen to align with most basic filter sets. However, the inclusion of more fluorochromes in your immunostainings adds complexity. For reliable imaging of tissues stained with up to seven colors (7-plex IHC), we offer a patented combination of filter sets—SpectraSplit® 7—which accommodates all the channels in the chart. SpectraSplit® 7 performs excellently with AZDyes and a variety of commercial dyes, such as AlexaFluor® dyes, FITC, TRITC, Cy3, Cy5, Cy7 dyes, and many of the OpalTM dyes.
|Channel||Traditional dyes||StreptaClick® dyes|
|S-Split Blue (375)||DAPI||AZDye™ 405|
|S-Split Cyan (435)||CF®430|
|S-Split Green (490)||FITC||AZDye™ 488|
|S-Split Orange (545)||Cy3/TRITC||AZDye™ 555,
Cyanine 3, Atto™ 542
|S-Split Red (590)||Texas Red||AZDye™ 594|
|S-Split Far-red (650)||Cy5/Cy5.5||AZDye™ 647,
|S-Split Infra-red (740)||Cy7|