GFAP Rabbit Polyclonal Antibody

• Catalog Number: orb500829
• Category: Antibodies
• Description: GFAP Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P
• Predicted Reactivity: Bovine, Canine, Porcine, Rabbit, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human GFAP (51-150/432aa)
• Concentration: 1mg/ml
• Dilution range: IHC-P=1:200-1000, IHC-F=1:200-1000, IF=1:200-800, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 48 kDa
• Target: GFAP
• UniProt ID: P14136
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Astrocyte Marker; FLJ45472; GFAP; Glial Fibrillary
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: RSC96 (blue). Primary Antibody: Rabbit Anti-GFAP antibody (orb500829), dilution: 1 µg 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 1X 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 (orb500829, 1 µg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1X 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 20000 events was performed.

Blank control: U87MG. Primary Antibody (green line): Rabbit Anti-GFAP antibody (orb500829), dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.

Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.

Paraformaldehyde-fixed, paraffin embedded (rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.

Paraformaldehyde-fixed, paraffin embedded (rat cerebellum), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat cerebellum), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (orb500829) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.

Sample: Optic nerve (Rat) cell Lysate at 40 ug, Primary: Anti-GFAP (orb500829) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution, Predicted band size: 48 kD, Observed band size: 53 kD.