SnapChip Akt pathway 11-plexProduct information
The Big Picture, in a SNAP
The SnapChip™ is perfectly suited to quickly and efficiently study the Akt pathway. The cross reaction-free, multiplex platform allows for a unique combination of assays, such as isoforms and phosphospecific antibodies, side-by-side.
Multiplex Sandwich Assays
Multiplexed sandwich immunoassay is a powerful technique to measure multiple protein concentrations simultaneously. Its wide adoption is hampered by inadequate antibody specificity which results in cross-reactivity and false positive signals (Fig. 1). This is a known weakness of multiplexed assays that is mitigated by months of assay optimization and validation, and despite considerable effort, it’s often impossible to combine related analytes in the same assay.
|Figure 1. Multiplexed assay. Cross-reaction occurs because the detection antibodies are mixed and applied as a “soup” to the array. Five scenarios of cross-reactivity are possible1. In this example, the signal on spot 2 is wrongly amplified by the binding of the blue protein to the red capture antibody.|
Parallex BioAssays Inc. substituted the “detection soup” by an array of detection nanodroplets and offers a new MICROARRAY-to-MICROARRAY approach to deliver the detection antibodies precisely to their cognate spots.2 As a result, a cross-reaction free chip harboring an assembly of parallelized assays, physically isolated from each other (Fig. 2).
Figure 2. SnapChipTM assay. The absence of mixing keeps the blue detection antibody from binding to the spot 2, thus leaving the wrong protein binding event with no consequence. Colocalization of capture and detection antibodies is an efficient approach to eliminate the cross-reaction.
1. Mix-&-Match – All your preferred assays on the same chip. No limitation.
- No more need to split your samples and run more than one assay, up to 500-plex (vs 5-10-plex).
- Off-the-shelves chips are available with unique target combinations!
2. Small Volume – Get the most of your precious samples. Always.
- Run an assay with as low as 20µL total.
3. Rapid assay development – All assays are optimized individually. That’s it
- Assemble multiplex in less than 3 weeks (vs 4-6 months)
4. Accurate Results - Despite rigorous assays validation, no one can predict cross-reaction in samples. We eliminated it.
- Reproduce the quality of a 96-well plate ELISA.
5. New opportunities - such as the measurement of concentrations, post-translational modifications and enzyme activities3… all at once.
- Ideal for cell signaling pathway activation studies
The assay process flow is highly similar to ELISA or regular multiplexed sandwich assay on planar microarray. Indeed, only the pipetting of the detection antibodies is replaced by a “SNAP” using the Snap Device (Fig. 3). The “detection soup” is eliminated as the detection antibodies are precisely delivered to their cognate spots during this microarray-to-microarray transfer3,4.
1. Pla-Roca, M. et al. Mol. Cell. Proteomics MCP 11, M111.011460 (2012).
2. Li, H., Bergeron, S. & Juncker, D. Anal. Chem. 84, 4776–4783 (2012).
3. Li, H., Munzar, J. D., Ng, A. & Juncker, D. Sci. Rep. 5, 11688 (2015).
4. Li, H., Bergeron, S, Larkin, H. & Juncker. JoVE (129), e56230 (2017)
The supporting documents available for this product can be downloaded below.