Fluorescently labeled secondary antibodies are needed to detect your primary sample (e.g. antibody in serum, purified antibody or protein). Strongly charged fluorescent dyes may interact with acidic and basic side chains of peptides. Therefore, we recommend the use of neutral dyes or dyes with few charged functional groups (e.g. DyLight680/DyLight800 or Cy3/Cy5), as well as pre-incubation with secondary antibodies to screen for background interactions.
We recommend the use of validated secondary antibodies from Rockland Immunochemicals, Inc., and offer additional support in in selecting the right secondary antibody and conjugate for the optimal detection of your samples. Please do not hesitate to talk to our scientists to know more about which Rockland secondary antibodies may be best suited for your project.
Product information
• | Application: DyLight680 and DyLight800 labelling for infrared scanners, or Cy3 and Cy5 labelling for compatible scanners (e.g., GenePix or Agilent) |
• | Recommended working dilution: 1:2000 - 1:5000 |
See also
Learn how to perform the assay
• | PEPperCHIP® Immunoassay Protocol [download pdf] [watch video] |
• | PEPperCHIP® Immunoassay Quickstart Guide [download pdf] |
Steps | Materials and equipment | We recommend |
Immunoassay | Incubation tray | PEPperCHIP® Incubation Tray* |
Orbital shaker | - | |
Blocking reagents | BSA, or Rockland Blocking Buffer* | |
Control antibodies | PEPperCHIP® Control Antibodies | |
Buffers | PBS, Tween 20 | |
Labelled secondary antibodies | Rockland secondary antibodies* | |
Data acquisition | Microarray scanner | Innopsys InnoScan® 710/710-IR* |
Data analysis | Fluorescence spot intensity quantification software | MAPIX Analyzer by Innopsys |
Spreadsheet software | - |
*used in-house by PEPperPRINT
<s>Take full control and perform PEPperCHIP® immunoassays at your own facility. Test multiple samples or multiple microarrays at once. Easy to assemble and fully reusable.</s>
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