Ordering Information
NeuroTox-2:High Content Analysis Kit for Gliosis
Description:
NeuroTox-2:High Content Analysis Kit for Gliosis
Product Overview:
Millipore’s HCS221 assay provides a complete solution for identifying and quantifying glial fibrillary acidic protein (GFAP) in cellular imaging studies. The reagents in the kit have been specifically optimized for HCS applications.
The assay is designed to enable visualization and quantitative detection of GFAP, allowing the characterization of agents that may induce or inhibit cellular events such as altered GFAP expression, reactive gliosis, neurotoxicity, astrocyte morphology and astrocyte differentiation. The nuclear dye (Hoechst 33342) may be used for measurements of cell number, DNA content and nuclear size. Additionally, the assay can be multiplexed with other probes, e.g., for signaling pathway studies, drug efficacy trials or in vitro toxicology applications.
The assay is immunofluorescence-based, and utilizes a high quality mouse monoclonal antibody which identifies GFAP in human, rodent, rabbit, bovine, porcine and avian cells. Additional species cross-reactivity is expected, but must be confirmed by the end user.
The assay is designed to enable visualization and quantitative detection of GFAP, allowing the characterization of agents that may induce or inhibit cellular events such as altered GFAP expression, reactive gliosis, neurotoxicity, astrocyte morphology and astrocyte differentiation. The nuclear dye (Hoechst 33342) may be used for measurements of cell number, DNA content and nuclear size. Additionally, the assay can be multiplexed with other probes, e.g., for signaling pathway studies, drug efficacy trials or in vitro toxicology applications.
The assay is immunofluorescence-based, and utilizes a high quality mouse monoclonal antibody which identifies GFAP in human, rodent, rabbit, bovine, porcine and avian cells. Additional species cross-reactivity is expected, but must be confirmed by the end user.
Background Information:
Glial fibrillary acidic protein (GFAP) as a biomarker of neurotoxicity amenable to High Content Screening
GFAP is an intermediate filament protein expressed predominantly in the astrocytes of the central nervous system (CNS). Astrocytes have an integral role in the maintenance of CNS homeostasis, and are also associated with both neuroprotection and neurodegradation when they are activated in response to toxic substances or disease states. Astrocytic activation (gliosis) leads to the upregulation of GFAP commonly accompanied by astrocyte proliferation and hypertrophy. This process of reactive gliosis has been proposed as an early marker of damage to the nervous system and can be detected morphologically or quantified by measurements of GFAP.
GFAP is an intermediate filament protein expressed predominantly in the astrocytes of the central nervous system (CNS). Astrocytes have an integral role in the maintenance of CNS homeostasis, and are also associated with both neuroprotection and neurodegradation when they are activated in response to toxic substances or disease states. Astrocytic activation (gliosis) leads to the upregulation of GFAP commonly accompanied by astrocyte proliferation and hypertrophy. This process of reactive gliosis has been proposed as an early marker of damage to the nervous system and can be detected morphologically or quantified by measurements of GFAP.
Key Applications:
- High Content Analysis
- Immunofluorescence
- in vitro Toxicology Assays
- Neuroscience
- Neurotoxicity
Application Notes:
Identification and quantification of glial fibrillary acidic protein (GFAP) in cellular imaging studies.
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.
Kit or Assay Type:
- High-Content Screening
- High-Content Analysis
Configuration:
Sufficient reagents for 5 X 96 well plates
Components:
- 1. Mouse Anti-GFAP HCS Primary Antibody, 100X
- 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. Acrylamide (Control Compound), 10X
- 8. Hydrogen Peroxide (Control Compound), 10X
- 9. K-252a (Control Compound), 250X
- 10. DMSO for Compound Serial Dilution
- 11. Compound Dilution Buffer
- 12. Plate Sealers
Storage Conditions:
Store kit components under the conditions indicated on the labels. Discard any remaining reagents after 6 months.
Detection Methods:
Fluorescent
Antibody Category:
Neuroscience
Materials Required but Not Delivered:
1. Sterile, tissue culture-treated black/clear bottom microplates suitable for High-Content Imaging.
2. Cell-type for assay, e.g., primary human or rodent astrocytes; astrocytoma or glioma cell lines.
3. Tissue culture instruments/supplies (including 37ºC incubator, growth media, flasks/plates, detachment buffer, etc.) for cell type of interest.
4. Sterile distilled or deionized water (dH2O) for Control Compound dilution, as appropriate.
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 and red ranges.
2. Cell-type for assay, e.g., primary human or rodent astrocytes; astrocytoma or glioma cell lines.
3. Tissue culture instruments/supplies (including 37ºC incubator, growth media, flasks/plates, detachment buffer, etc.) for cell type of interest.
4. Sterile distilled or deionized water (dH2O) for Control Compound dilution, as appropriate.
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 and red ranges.