bs-0199R [Primary Antibody]
Rabbit  Anti-GFAP Polyclonal Antibody
www.sanhaoinfo.com
sales@ag88.com.com.cn
support@ag88.com.com.cn
400-901-9800
DATASHEET

Host: Rabbit

Target Protein: GFAP

IR: Immunogen Range:51-150432

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 2670

Swiss Prot: P14136

Source: KLH conjugated synthetic peptide derived from human GFAP:51-150432 

Purification: affinity purified by Protein A

Storage: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Background: 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. [provided by RefSeq, Oct 2008]

Size: 200ul

Concentration: 1mg/ml

Applications: WB(1:500-2000)
ELISA(1:500-1000)
IHC-P(1:200-1000)
IHC-F(1:200-1000)
Flow-Cyt(1μg/Test)
ICC(1:100-500)
IF(1:200-800)

Cross Reactive Species: Human
Mouse
Rat
Dog
Pig
Cow
Rabbit
Sheep
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample:
Cerebellum (Rat) Lysate at 40 ug
Cerebellum (Mouse) Lysate at 40 ug
Eye (Mouse) Lysate at 40 ug
Primary: Anti-GFAP (bs-0199R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 48 kD
Sample:Optic nerve (Rat)cell Lysate at 40 ug
Primary: Anti-GFAP(bs-0199R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 53 kD
Sample: U251 Cell Lysate at 40 ug
Primary: Anti- GFAP (bs-0199R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution
Predicted band size: 48 kD
Observed band size: 50 kD
Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-GFAP Polyclonal Antibody, Unconjugated(bs-0199R) 1:400, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-GFAP Polyclonal Antibody, Unconjugated(bs-0199R) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Paraformaldehyde-fixed, paraffin embedded (Human brain glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by a conjugated secondary (bs-0295G-Cy3) at [1:500] for 90 minutes and DAPI staining of the nuclei.
Tissue/cell: rat brain tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-GFAP Polyclonal Antibody, Unconjugated(bs-0199R) 1:400, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
Blank control: RSC96(blue).
Primary Antibody:Rabbit Anti- GFAP antibody(bs-0199R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (bs-0199R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.
 
PRODUCT SPECIFIC PUBLICATIONS