PathScan® Antibody Arrays
Our scientists are at the bench daily to produce and validate our antibodies, so they have hands-on experience and knowledge of each antibody’s performance.
PathScan® Antibody Array Kits from Cell Signaling Technology (CST) are slide-based assays using the sandwich ELISA principle, and are designed to investigate signal transduction occurring through key pathway nodes in a multiplex format. Two slides are provided in each kit; each slide contains arrays spotted with multiple antibodies. Captured proteins are subsequently detected using a detection antibody cocktail. Positive and negative controls are also included on each array.
Kits are available for both chemiluminescent and fluorescent detection; chemiluminescent array detection is adaptable to film, making any additional equipment unnecessary. Fluorescent readouts can be performed using a fluorescent imager.
- Arrays are produced and optimized in-house with high-quality and thoroughly validated antibodies, providing you with the greatest possible specificity and sensitivity.
- Arrays allow the analysis of multiple proteins with small lysate volume, saving valuable time and reagents.
- Chemiluminescent detection allows convenient and easy detection without specialized equipment.
- Technical Support is provided by CST’s in-house Molecular Assay Group who develops and produces the products and knows them best.
Search our extensive line of PathScan® Antibody Array Kits for the pathways and nodes most relevant to your research.
What is the difference between the fluorescent and chemiluminescent kit? Which one would you recommend?
This depends on the desired outcome of the experiment and the availability of imagers to capture the data. Our fluorescent detection kits provide higher sensitivity and ease of quantitation. These kits require an imager capable of detecting the fluorescent signal (see next question). For more qualitative studies, our chemiluminescent kits provide a convenient readout. Our chemiluminescent kits are designed so that any lab that is equipped to perform western blots can also use our antibody array products.
For our chemiluminescent detection kits you can expose the array to film. A digital camera based imager may also be used if it has sufficient resolution.
For our fluorescent detection kits, you will need an imager capable of exciting at 680nm and detecting at 700nm, such as, the LI-COR® Odyssey®, Typhoon™ FLA 9000 or INNOPSYS Innoscan® 710-IR. Alternatively, Streptavidin linked Alexa Fluor® 647 (user supplied) can be used in place of the streptavidin-linked DyLight™ 680 secondary supplied with the kit. Streptavidin-linked Alexa-Fluor® 647 (excitation 594nm, emission 633nm) can be detected using a number of different scanners including: The Axon® GenePix® 4100A Microarray Scanner, INNOPSYS Innoscan® microarray scanners, the Tecan® PowerScanner™, the Roche NimbleGen™ MS 200 microarray scanner, Alpha Innotech AlphaScan microarray scanner, or the Agilent SureScan microarray scanner.
We recommend the following settings as a good starting point:
Setup: Grid Array
Controls: Resolution, 21; Quality, High; Focus offset, 0.0 mm
Channels: Check the 700 box, deselect the 800 box, and deselect Auto Channel
Which detection method (chemiluminescent or fluorescent) gives a rapidly saturating signal?
Chemiluminescent. Typically, we do not expose the array to film for more than 15 seconds. Fluorescent-based detection uses a rapid surface scan where the degree of saturation is modified by changing either the PMT (photomultiplier tube) setting or laser power.
Our glass slides are 25 mm x 75 mm (1 in x 3 in).
Although we have not tested this ourselves, a number of our customers have successfully examined tissue samples with our array kits. We recommend using the kit supplied cell lysis buffer (product #9803 or #7018) at a 2X concentration when working with tissues. This will increase the concentration of protease and phosphatase inhibitors present during lysis.
We recommend 0.5 mg/ml as a good starting point. The majority of our in-house quality control experiments are performed using lysates at 0.5 mg/ml of total protein, though some range from 0.1 to 1.0 mg/ml.
You can use the lysates noted in the figures of our datasheets as a positive control. We do not have one cell lysate that expresses all the targets represented in the array.
We do not recommend using part of the slide, as this may harm the remaining wells. We do supply two slides per kit so you can perform two separate experiments if desired.
No. These spots should be used to ensure the detection steps of the assay were performed correctly and to determine the orientation of the slide.