SKU PA1239
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications IF, IHC, WB

Overview

Product Name Anti-GFAP Antibody
SKU/Catalog Number PA1239
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Glial fibrillary acidic protein(GFAP) detection. Tested with WB, IHC-P, IF in Human;Mouse;Rat.
Cite This Product Anti-GFAP Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1239)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence at the C-terminus of human GFAP(417-432aa DGEVIKESKQEHKDVM), identical to the related rat sequence, and different from the related mouse sequence by two amino acids.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Rat, Mouse, By Heat
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunofluorescence, 2μg/ml, Mouse

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).

Images And Assay Conditions

/antibody/pa1239 1 WB anti gfap antibody.jpg

Figure 1. Western blot analysis of GFAP using anti- GFAP antibody (PA1239).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Rat Brain Tissue Lysate,
Lane 2: Mouse Brain Tissue Lysate,
Lane 3: U87 Whole Cell Lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- GFAP antigen affinity purified polyclonal antibody (Catalog # PA1239) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GFAP at approximately 49 KD. The expected band size for GFAP is at 49 KD.

/antibody/pa1239 2 IHC anti gfap antibody.jpg

Figure 2. IHC analysis of GFAP using anti- GFAP antibody (PA1239).
GFAP was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- GFAP Antibody (PA1239) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

/p/a/pa1239 3_1.jpg

Figure 3. IF analysis of GFAP using anti- GFAP antibody (PA1239).
GFAP was detected in paraffin-embedded section of mouse brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- GFAP Antibody (PA1239) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P14136
Gene Name GFAP
Protein Name Glial fibrillary acidic protein
Tissue Specificity Expressed in cells lacking fibronectin. .
Alternative Names Glial fibrillary acidic protein;GFAP;GFAP;
Subcellular Localization Cytoplasm . Associated with intermediate filaments.
Molecular Weight 49880 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein 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.
Research Areas Cell Adhesion Proteins, Cell Type Marker, Cell Type Markers, Cytoskeleton, Cytoskeleton / Ecm, Intermediate Filaments, Neural Stem Cells, Neuroscience, Signal Transduction, Stem Cells, Tags & Cell Markers

*You can search these to find other products in these research areas.
Background Glial fibrillary acidic protein(GFAP) is an intermediate-filament(IF) protein that is highly specific for cells of astroglial lineage, although its tissue-specific role is speculative. GFAP has been located in rat kidney glomeruli and peritubular fibroblasts, leydig cells of testis, skin keratinocytes, osteocytes of bones, chondrocytes of epiglottis, bronchus, and stellate-shaped cells of the pancreas and liver. Its expression is essential for normal white matter architecture and blood-brain barrier integrity, and its absence leads to late-onset CNS dysmyelination. GFAP has also been shown to play a role in mitosis by adjusting the filament network present in the cell. During mitosis, there is an increase in the amount of phosphorylated GFAP, and a movement of this modified protein to the cleavage furrow.

Other Recommended Resources

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Order Product (PA1239)

Promotion:

Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
Option Price
30ug sample size $99
100ug $280
100ug+Free HRP Secondary BA1054 $280
100ug+Free Biotin Secondary BA1003 $280

USD $280

Ships in 7-10 business days.

Troubleshooting

Download troubleshooting handbooks for IHC, Western blot and ELISA for FREE.

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Boster Guarantee

Guaranteed product quality

We promise all of our products perform as described in datasheets.

Customer Reviews

Review by
Rating:

Source: Biocompare.com

"This Antibody Works Perfectly!"

--Maria Teresa Dell'Anno, NEUROSCIENCE, YALE UNIVERSITY, POSTDOC

Applications Immunohistochemistry
Sample Spinal cord
Detection Confocal microscope

"I used it for IHC on frozen sections at a dilution of 1:500. It did not need several trials to optimize the protocol. No bad things overall. I will purchase it again."

(Posted on 8/23/2016)
Write a review for PA1239

Publications

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Reversion of malignant phenotypes of human glioblastoma cells by?-elemene through?-catenin-mediated regulation of stemness-, differentiation- and epithelial-to-mesenchymal transition-related molecules
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Establishment of immortalized Schwann cells derived from rat embryo dorsal root ganglia
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Wang R, Zhang X, Zhang J, Fan Y, Shen Y, Hu W, Chen Z. Plos One. 2012;7(5):E37574. Doi: 10.1371/Journal.Pone.0037574. Epub 2012 May 22. Oxygen-Glucose Deprivation Induced Glial Scar-Like Change In Astrocytes.
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Customer Q&As

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands

3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.

4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to gfap antibody, astrocyte antibody, glial fibrillary acidic protein antibody