GFAP ELISA

Analytes Available	Species Reactivity	Key Applications	Detection Methods
GFAP	M, R, H	ELISA	Chromogenic

Analytes Available

Species Reactivity

Key Applications

Detection Methods

GFAP

M, R, H

ELISA

Chromogenic

Description
Catalogue Number	NS830
Brand Family	Chemicon®
Trade Name	Chemicon
Description	GFAP ELISA Kit
Overview	Background Glial Fibrillary Acidic Protein (GFAP) is a class-III intermediate filament uniquely and ubiquitously expressed as part of the structural network in astrocytic glial cells. This specific expression makes GFAP ideal as a broad astrocytic marker in vertebrates. Astrocytes play a variety of key roles in supporting, guiding, nurturing, and signaling neuronal architecture and activity in a highly interdependent manner. Current understanding of CNS structure suggests that there are at least as many glial cells as the estimated 100 billion neurons, but perhaps upwards of 100 times more. Thus, the health of both neurons and astrocytes is often affected in disease and trauma. GFAP is proving to be a useful biomarker in the detection of neural degeneration. Although defects in GFAP have been reported to cause certain rare conditions, such as Alexander’s disease (Brenner et al., 2001), the observed interconnected physiology of neuronal and glial cells has enabled the use of GFAP as a neurodegeneration detection gauge. This relationship is being exploited by researchers and clinicians measuring a variety of neurological issues from cancer to trauma. GFAP has been used as a biomarker in the early detection of chemically induced cancer (Capo et al., 1997) and classical glioblastomas (Jung et al., 2007). Elevated levels of GFAP have been reported in traumatic brain injury (TBI) and appear to directly correlate with the severity of injury and brain tissue oxygenation in combat casualty care research (CCR, 2008) as well as civilian accidents (Mondello et al., 2010). GFAP has also been identified as a good biomarker in studies of toxin induced acute brain injury (Liao et al., 2008), in differentiation of hemorrhagic vs. ischemic strokes (Foerch et al., 2006), as well as in cardiac arrest (Kaneko et al., 2009). Millipore now offers an ELISA assay for sensitive measurement of GFAP protein in tissue homogenates and biological fluids (serum, plasma, CSF, and serum free media) from human, mouse, and rat. Principals of Procedure This assay is a Sandwich ELISA based sequentially on: 1) capture of soluble GFAP protein to the wells of a microtiter plate coated by a pre-titered amount of anti-GFAP monoclonal antibody; 2) removal of unbound material by washing; 3) binding of biotinylated anti-GFAP monoclonal antibody to the previously captured GFAP protein; 4) removal of unbound material by washing; 5) binding of streptavidin conjugated horseradish peroxidase to the biotinylated antibodies; 6) removal of excess or unbound enzyme conjugates by washing; and 7) quantification of immobilized antibody-enzyme complexes by monitoring horseradish peroxidase activity in the presence of the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). Enzyme activity is measured spectrophotometrically by recording absorbance readings at 450 nm. Increases in absorbency are directly proportional to the amount of GFAP protein captured from the sample. Therefore the protein concentration of unknowns can be derived by interpolation using a standard curve generated in the same assay with reference standards of known concentrations of GFAP.
Alternate Names	GFAP Glial Fibrillary Acidic Protein ELISA
Materials Required but Not Delivered	1. Absorbent paper or cloth 2. Conventional, multi-channel, or repeating pipettes 3. Reagent reservoirs 4. Orbital microtiter plate shaker/washer (optional) 5. Graduated serological pipettes 6. Microtiter plate reader capable of reading absorbency at 450 nm 7. Graphing software for plotting data or graph paper for manual plotting of data 8. Microfuge tubes for standard and sample dilutions 9. Mechanical vortex 10. Milli-Q™ Water

Description

Catalogue Number

NS830

Brand Family

Chemicon®

Trade Name

Chemicon

Description

GFAP ELISA Kit

Overview

Background
Glial Fibrillary Acidic Protein (GFAP) is a class-III intermediate filament uniquely and ubiquitously expressed as part of the structural network in astrocytic glial cells. This specific expression makes GFAP ideal as a broad astrocytic marker in vertebrates. Astrocytes play a variety of key roles in supporting, guiding, nurturing, and signaling neuronal architecture and activity in a highly interdependent manner. Current understanding of CNS structure suggests that there are at least as many glial cells as the estimated 100 billion neurons, but perhaps upwards of 100 times more. Thus,
the health of both neurons and astrocytes is often affected in disease and trauma. GFAP is proving to be a useful biomarker in the detection of neural degeneration. Although defects in GFAP have been reported to cause certain rare conditions, such as Alexander’s disease (Brenner et al., 2001), the observed interconnected physiology of neuronal and glial cells has enabled the use of GFAP as a neurodegeneration detection gauge. This relationship is being exploited by researchers and clinicians measuring a variety of neurological issues from cancer to trauma. GFAP has been used as a biomarker in the early detection of chemically induced cancer (Capo et al., 1997) and classical glioblastomas (Jung et al., 2007). Elevated levels of GFAP have been reported in traumatic brain injury (TBI) and appear to directly correlate with the severity of injury and brain tissue oxygenation in combat casualty care research (CCR, 2008) as well as civilian accidents (Mondello et al., 2010). GFAP has also been identified as a good biomarker in studies of toxin induced acute brain injury (Liao et al., 2008), in differentiation of hemorrhagic vs. ischemic strokes (Foerch et al., 2006), as well as in cardiac arrest (Kaneko et al., 2009). Millipore now offers an ELISA assay for sensitive measurement of GFAP protein in tissue homogenates and biological fluids (serum, plasma, CSF, and serum free media) from human, mouse, and rat.

Principals of Procedure
This assay is a Sandwich ELISA based sequentially on: 1) capture of soluble GFAP protein to the wells of a microtiter plate coated by a pre-titered amount of anti-GFAP monoclonal antibody; 2) removal of unbound material by washing; 3) binding of biotinylated anti-GFAP monoclonal antibody to
the previously captured GFAP protein; 4) removal of unbound material by washing; 5) binding of streptavidin conjugated horseradish peroxidase to the biotinylated antibodies; 6) removal of excess or unbound enzyme conjugates by washing; and 7) quantification of immobilized antibody-enzyme
complexes by monitoring horseradish peroxidase activity in the presence of the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). Enzyme activity is measured spectrophotometrically by recording absorbance readings at 450 nm. Increases in absorbency are directly proportional to the amount of
GFAP protein captured from the sample. Therefore the protein concentration of unknowns can be derived by interpolation using a standard curve generated in the same assay with reference standards of known concentrations of GFAP.

Alternate Names

GFAP
Glial Fibrillary Acidic Protein ELISA

Materials Required but Not Delivered

1. Absorbent paper or cloth
2. Conventional, multi-channel, or repeating pipettes
3. Reagent reservoirs
4. Orbital microtiter plate shaker/washer (optional)
5. Graduated serological pipettes
6. Microtiter plate reader capable of reading absorbency at 450 nm
7. Graphing software for plotting data or graph paper for manual plotting of data
8. Microfuge tubes for standard and sample dilutions
9. Mechanical vortex
10. Milli-Q™ Water

Product Information
Components	96-Well Anti-GFAP Coated Plate (CS206352): 1 Plate 1X Assay Buffer (CS206357): 40 mL 10X Wash Buffer (CS206360): 50 mL GFAP Protein Standard (CS206361): 1 Vial GFAP Quality Control #1 (CS206351): 1 Vial GFAP Quality Control #2 (CS206475): 1 Vial Biotinylated Anti-GFAP Detection Antibody (CS206356): 12 mL Enzyme Solution (CS206358): 12 mL TMB Solution (CS202682): 10 mL Stop Solution (60193): 12 mL Plate Sealers (CS200443): 10 Sheets
Detection method	Chromogenic

Product Information

Components

96-Well Anti-GFAP Coated Plate (CS206352): 1 Plate
1X Assay Buffer (CS206357): 40 mL
10X Wash Buffer (CS206360): 50 mL
GFAP Protein Standard (CS206361): 1 Vial
GFAP Quality Control #1 (CS206351): 1 Vial
GFAP Quality Control #2 (CS206475): 1 Vial
Biotinylated Anti-GFAP Detection Antibody (CS206356): 12 mL
Enzyme Solution (CS206358): 12 mL
TMB Solution (CS202682): 10 mL
Stop Solution (60193): 12 mL
Plate Sealers (CS200443): 10 Sheets

Detection method

Chromogenic

Applications
Key Applications	ELISA

Applications

Key Applications

ELISA

Biological Information
Species Reactivity	Mouse Rat Human
Analytes Available	GFAP
Entrez Gene Number	NM_002055.2
Entrez Gene Summary	This gene encodes one of the major intermediate filament proteins of mature astrocytes. It is used as a marker to distinguish astrocytes from other glial cells during development. Mutations in this gene cause Alexander disease, a rare disorder of astrocytes in the central nervous system. Alternative splicing results in multiple transcript variants encoding distinct isoforms.
Gene Symbol	GFAP
UniProt Number	P14136
UniProt Summary	FUNCTION: GFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells. SUBCELLULAR LOCATION: Cytoplasm. Note: Associated with intermediate filaments. TISSUE SPECIFICITY: Expressed in cells lacking fibronectin. PTM: Phosphorylated by PKN1. SIMILARITY: Belongs to the intermediate filament family.

Biological Information

Species Reactivity

Mouse
Rat
Human

Analytes Available

GFAP

Entrez Gene Number

NM_002055.2

Entrez Gene Summary

This gene encodes one of the major intermediate filament proteins of mature astrocytes. It is used as a marker to distinguish astrocytes from other glial cells during development. Mutations in this gene cause Alexander disease, a rare disorder of astrocytes in the central nervous system. Alternative splicing results in multiple transcript variants encoding distinct isoforms.

Gene Symbol

GFAP

UniProt Number

P14136

UniProt Summary

FUNCTION: GFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells.
SUBCELLULAR LOCATION: Cytoplasm. Note: Associated with intermediate filaments.
TISSUE SPECIFICITY: Expressed in cells lacking fibronectin.
PTM: Phosphorylated by PKN1.
SIMILARITY: Belongs to the intermediate filament family.

Physicochemical Information
Sensitivity	1.5 ng/mL
Standard Curve Range	1.5 to 100 ng/mL

Physicochemical Information

Sensitivity

1.5 ng/mL

Standard Curve Range

1.5 to 100 ng/mL

Product Usage Statements
Usage Statement	For research use only. Not for use in diagnostic procedures.

Product Usage Statements

Usage Statement

For research use only. Not for use in diagnostic procedures.

Storage and Shipping Information
Storage Conditions	Storage: The GFAP ELISA kit is shipped on ice and should be stored at 2-8°C upon arrival. Stability: Kit components are stable for a minimum of 6 months from the date of receipt if stored and handled as described above.

Storage and Shipping Information

Storage Conditions

Storage: The GFAP ELISA kit is shipped on ice and should be stored at 2-8°C upon arrival.
Stability: Kit components are stable for a minimum of 6 months from the date of receipt if stored and
handled as described above.

Packaging Information
Material Size	1 kit
Material Package	96 wells

Packaging Information

Material Size

1 kit

Material Package

96 wells

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

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