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Alexa Fluor® 647 anti-GFAP Antibody

Pricing & Availability
Clone
2E1.E9 (See other available formats)
Regulatory Status
RUO
Other Names
Glial fibrillary acid protein
Isotype
Mouse IgG2b
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Product Citations
publications
a-2E1-E9_A647_GFAP_Antibody_1_022620
C57BL/6 mouse frozen cerebellum section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS for 1 hour at room temperature. Then the section was stained with 2.5 µg/mL of GFAP (clone 2E1.E9) Alexa Fluor® 647 (magenta) overnight at 4°C. Nuclei were counterstained with DAPI (green). The image was captured by 10X objective. Scale Bar: 50 µm
  • a-2E1-E9_A647_GFAP_Antibody_1_022620
    C57BL/6 mouse frozen cerebellum section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS for 1 hour at room temperature. Then the section was stained with 2.5 µg/mL of GFAP (clone 2E1.E9) Alexa Fluor® 647 (magenta) overnight at 4°C. Nuclei were counterstained with DAPI (green). The image was captured by 10X objective. Scale Bar: 50 µm
  • b-2E1dotE9_A647_GFAP_Antibody_IHC_2_091117
    Human paraffin-embedded cerebellum tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate Buffered 1X (1.0M, pH6.0) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10µg/mL of Alexa Fluor® 647 anti-GFAP Antibody (Clone 2E1.E9, red) and Alexa Fluor® 594 anti-Tubulin Beta 3 (TUBB3) Antibody (Clone AA10, green) antibody overnight at 4°C. Nuclei were counterstained with DAPI (blue). The image was scanned with a 10X objective and stitched with MetaMorph® software.
  • c-2E1dotE9_A647_GFAP_Antibody_IHC_3_091117
    Human paraffin-embedded cerebellum tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate Buffered 1X (1.0M, pH6.0) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10µg/mL of Alexa Fluor® 647 anti-GFAP Antibody (Clone 2E1.E9, red) and Alexa Fluor® 594 anti-Tubulin Beta 3 (TUBB3) Antibody (Clone AA10, magenta) antibody overnight at 4°C. Nuclei were counterstained with DAPI (green). The image was scanned with a 10X objective and stitched with MetaMorph® software.
Compare all formats See Alexa Fluor® 647 spectral data
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644706 100 µg 323 CHF
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Description

GFAP is expressed exclusively in astrocytes in the central nervous system. The protein is a member of the intermediate filament family of proteins which form networks providing support and strength to cells. This antibody does not cross-react with other intermediate filaments such as vimentin, neurofilament proteins, desmin and others. More than 50 GFAP mutations have been identified to be associated with the Alexander disease.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Bovine spinal cord homogenate
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

IHC-F - Quality tested
IHC-P - Verified
SB - Community verified
Recommended Usage

Each lot of this antibody is quality control tested by immunohistochemical staining on frozen tissue sections. For immunohistochemistry, a concentration range of 1.0 - 5.0 µg/mL is suggested. For immunohistochemical staining on formalin-fixed paraffin-embedded tissue sections, the suggested use of this reagent is 5.0 - 10 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) and IHC-P (Immunohistochemistry - formalin-fixed paraffin-embedded tissues) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

(PubMed link indicates BioLegend citation)
  1. McLendon RE and Bigner DD. 1994. Brain Pathol. 4:221.
  2. Liu W, et al. 2011. Proteomics 11:3556. (FC) PubMed
Product Citations
  1. Khan NZ, et al. 2021. Brain Behav Immun. 92:165. PubMed
  2. Tyler CR, et al. 2018. Toxicol Sci. 163:123. PubMed
  3. Rocktäschel P, et al. 2019. Epilepsy Behav. 101:106581. PubMed
RRID
AB_2566110 (BioLegend Cat. No. 644706)

Antigen Details

Structure
Around 50 kD
Distribution

Only expressed in astrocytes in the central nervous system.

Function
Together with other proteins to form intermediate filaments which supports astroglial cells. Astroglial cells support and nourish cells in the brain and spinal cord. If brain cells are injured through trauma or disease, astroglial cells react by rapidly prodcing more GFAP protein.
Cell Type
Astrocytes
Biology Area
Cell Biology, Neuroscience, Neuroscience Cell Markers
Molecular Family
Intermediate Filaments
Gene ID
2670 View all products for this Gene ID
UniProt
View information about GFAP on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

View All GFAP Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-GFAP 2E1.E9 WB,ICC,IHC-F,IHC-P,FC
Alexa Fluor® 488 anti-GFAP 2E1.E9 IHC-F,ICC,IHC-P,SB
Alexa Fluor® 647 anti-GFAP 2E1.E9 IHC-F,IHC-P,SB
Alexa Fluor® 594 anti-GFAP 2E1.E9 IHC-F,IHC-P,SB
Brilliant Violet 421™ anti-GFAP 2E1.E9 IHC-F,ICC,IHC-P
Go To Top Version: 3    Revision Date: 01.24.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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