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

StreptaClick®-Color 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.

Exceptional Fluorochromes

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.

Imaging

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,
Cyanine 5
S-Split Infra-red (740) Cy7

SpectraSplit® 7

SpectraSplit® 7 are patented filter sets that empower fluorescence microscopes with seven color channels.  The channels are distributed over four filter sets and specifically separate the most commonly used fluorochromes in immunofluorescence (see table below). Hence, you can immunostain tissues with up to seven fluorochromes and still visualize each fluorochrome in the sample with SpectraSplit® 7 without bleedthrough. No additional software and no computational spectral unmixing are required. SpectraSplit® 7 can easily be installed in both standard and scanning fluorescence microscopes, and can be run with different popular light sources. See the description below for more information. Request a Quote

StreptaClick® Color

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StreptaClick® Color labels antibodies with fluorochromes, mimicking directly conjugated antibodies. StreptaClick® Color provides a rapid, robust, and flexible method for attaching fluorochromes to biotinylated antibodies. Once labeled with StreptaClick®, the antibody is stable and can be used in the same manner as antibodies that are directly conjugated with fluorochromes. The click reaction is highly efficient and can be carried out at any volume, concentration, or buffer. StreptaClick® Color is available in four colors: AZDye™ 488, AZDye™ 594, AZDye™ 647, and Atto™ 542. The AZDyes™ are structurally identical to the Alexa Fluor® dyes, and Atto 542 is a particularly bright dye with a spectral similarity to Cy®3. The StreptaClick® 2-Color Kit contains the Labeling Reagents for StreptaClick® Color 488 and StreptaClick® Color 647, along with Biotin Block Buffer. The StreptaClick® 2-Color Kit is suitable for multiplex IHC and Flow Cytometry. The StreptaClick® 4-Color Kit contains the Labeling Reagents for StreptaClick® Color 488, StreptaClick® Color 542, StreptaClick® Color 594, and StreptaClick® Color 647, along with Biotin Block Buffer. The StreptaClick® 4-Color Kit is suitable for multiplex IHC. Each color contains 500 μl of StreptaClick® antibody labeling reagent, capable of labeling 50 μg of biotinylated antibody, sufficient for approximately 50 slides. Select your StreptaClick® Color Kit: