High Content Screening Toolkit
For Use With Mouse Primary Antibodies
Description:
High Content Screening Toolkit
For Use With Mouse Primary Antibodies
For Use With Mouse Primary Antibodies
Product Overview:
Millipore’s HCS215 High Content Screening Toolkit provides a set of validated reagents to enable the end-user to create a reproducible HCS assay using any mouse primary antibody of their choice. The superior reagents provided with this kit allow the user to standardize their HCS assays, minimize assay-to-assay variability, and reproducibly generate images with a high signal-to-background ratio.
Background Information:
Successful HCS assays rely on high quality reagents. With the commercial availability of thousands of immunoreagents and fluorescent probes, large numbers of fixed-endpoint HCS assays are possible. However, incompatibility of reagents when integrated into a single assay can lead to a significant drop-off in assay performance. Immunoreagents for HCS assays carry special requirements. Strong antigen affinity is required, minimal non-specific binding must be observed, interactions between multiple primary or secondary immunoreagents must be minimized, and the signal to background ratio must be sufficient to ensure an adequate screening window. Additionally, to enable scale up of High Content Screening assays, the sample preparation protocol must be highly reproducible, and the reagents must exhibit minimal assay-to-assay variability.
Key Applications:
in vitro Toxicology Assays
Application Notes:
Cell Cycle
Species Reactivity:
Mouse
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Therapeutic Areas:
- Cardiovascular Disease
- Metabolic Disorders
Host Cells:
HeLa cell line
Materials Required but Not Delivered:
1. Rabbit primary antibody of choice.
2. Sterile, tissue culture-treated black/clear bottom microplates suitable for High-Content Imaging.
3. Cell type for assay.
4. Tissue culture instruments/supplies (including 37ºC incubator, growth media, flasks/plates, detachment buffer, etc.) for cell type of interest.
5. HCS imaging/analysis system, e.g., GE Healthcare IN Cell Analyzer 1000 with Investigator software. Imaging system must be equipped with beam-splitters and filters capable of reading emission spectra in the blue, green or red ranges. Example filter ranges are shown in Table 2 below.
2. Sterile, tissue culture-treated black/clear bottom microplates suitable for High-Content Imaging.
3. Cell type for assay.
4. Tissue culture instruments/supplies (including 37ºC incubator, growth media, flasks/plates, detachment buffer, etc.) for cell type of interest.
5. HCS imaging/analysis system, e.g., GE Healthcare IN Cell Analyzer 1000 with Investigator software. Imaging system must be equipped with beam-splitters and filters capable of reading emission spectra in the blue, green or red ranges. Example filter ranges are shown in Table 2 below.
Components:
- 1. HCS Secondary Antibody (donkey anti-mouse IgG, FITC conjugate), 200X
- 2. HCS Secondary Antibody (donkey anti-mouse IgG, Cy3 conjugate), 200X
- 3. Hoechst HCS Nuclear Stain, 200X
- 4. HCS Fixation Solution with Phenol Red, 2X
- 5. HCS Immunofluorescence Buffer, 1X
- 6. HCS Wash Buffer, 1X
- 7. Plate Sealers
Kit Type:
High-Content Screening
Configuration:
Sufficient reagents for 5 X 96 well plates
Detection Methods:
Fluorescent
Kit or Assay Type:
High-Content Screening
Storage Conditions:
When stored under the conditions indicated on the labels, the kit components are stable up to the expiration date. HCS Fixation Solution, HCS Immunofluorescence Buffer, HCS Wash Buffer and Plate Sealers should be stored at 2-8ºC. HCS Secondary Antibodies and Hoechst HCS Nuclear Stain should be stored at -20ºC, avoiding repeated freeze/thaw cycles. (Note: If kit is expected to be used for multiple experiments, rather than a single use, thaw antibodies and nuclear stain and dispense into appropriately sized aliquots. Store aliquots at -20ºC.)
Discard any remaining reagents after the expiration date.
Discard any remaining reagents after the expiration date.