GFAP Rabbit Polyclonal Antibody

• Catalog Number: orb221420
• Category: Antibodies
• Description: GFAP Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, WB
• Predicted Reactivity: Bovine, Gallus, Porcine, Rabbit, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human GFAP (341-432/432aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, Flow-Cyt=3μg/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; FLJ45472; GFAP; Glial Fibrillary Acidic
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Maione, Francesco et al. Down regulation of pro-inflammatory pathways by tanshinone IIA and cryptotanshinone in a non-genetic mouse model of Alzheimer's disease Pharmacol Res, 129, 482-490 (2018)

PubMed: 29158049

Sample: Cerebrum (Mouse) Lysate at 40 ug, Primary: Anti-GFAP (orb221420) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 48 kD, Observed band size: 48 kD.

Sample: Cerebrum (Rat) Lysate at 40 ug, Primary: Anti-GFAP (orb221420) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 48 kD, Observed band size: 48 kD.

Tissue/Cell: SH-SY5Y cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (GFAP) polyclonal Antibody, Unconjugated (orb221420) 1:100, 90 minutes at 37°C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Blank control (blue line): Hela (fixed with 80% methanol (5 min at -20°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature). Primary Antibody (green line): Rabbit Anti-GFAP antibody (orb221420), dilution: 3 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, dilution: 1 µg/Test.

GFAP was immunoprecipitated from human hela cells lysate with orb221420 at 1/150 dilution. Western blot was performed from the immunoprecipitate using protein A/G beads. HRP Conjugated Mouse anti-Rabbit IgG (Light Chain specific) was used as secondary antibody at 1:5000 dilution. Lane 1: human hela cells lysate 10 µg (Input). Lane 2: orb221420 IP in human hela cells lysate. Lane 3: native rabbit IgG IP in human hela cells lysate (negative control). Secondary, All lanes: Mouse anti-Rabbit IgG (Light Chain specific), HRP Conjugated, 1:5000.