When selecting antibodies for the Teton Custom Add-On Protein Panel Assembly Kit, consider the following guidance for best performance.
What are the requirements for antibody selection and validation?
All antibodies selected or provided by the customer for use with the Teton Custom Add-On Protein Panel must be rabbit-derived, with monoclonal antibodies preferred to ensure optimal performance. At a minimum, antibodies should be validated for western blot (WB). When available, we recommend selecting antibodies that have also been validated for immunocytochemistry/immunofluorescence (ICC/IF) and flow cytometry, as those with multiple validation methods tend to perform more reliably with the Teton assay.
To ensure consistency and data quality, each antibody should be confirmed using an orthogonal method—typically western blot or immunofluorescence.
Can custom antibodies be used for phosphorylated protein detection?
The Teton Custom Add-On Protein Panel Assembly Kit works well with antibodies targeting both phosphorylated and unphosphorylated proteins. When considering phosphorylated targets, make sure that your antibody is specific to the phosphorylated site and/or state that you are looking for.
When is the Diversity Spike-In required?
If you are preparing a panel with fewer than or equal to 24 proteins, either prepare multiple wells per target to exceed 24 total barcodes OR use a Diversity Spike-In (830-00043). This will ensure that we have enough nucleotide diversity when sequencing the barcodes to generate high quality base calls.
Can I re-use the Teton Custom Add-On Protein Panel Plate if I use fewer than 88 barcodes?
No, the Teton Custom Add-On Protein Panel Plate is single use.
Can I utilize multiple barcodes for a single protein target to increase my sensitivity?
If you are selecting fewer than 88 antibodies for your panel, you may consider using multiple wells for a single antibody target. In doing this, you may improve the total number of counts per protein target, increasing sensitivity.