Osteopontin Rabbit Polyclonal Antibody

• Catalog Number: orb500935
• Category: Antibodies
• Description: Osteopontin Rabbit Polyclonal Antibody
• Target: SPP1
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Rat
• Predicted Reactivity: Human
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Immunogen: KLH conjugated synthetic peptide derived from rat osteopontin (201-314/314aa)
• UniProt ID: P08721
• MW: 34 kDa
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:500-2000, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test
• Antibody Type: Primary Antibody
• 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.
• Alternative names: BNSP; Bone sialoprotein 1; BSP I; BSPI; Early T ly
• Note: For research use only

Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-Osteopontin antibody (orb500935), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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.

Lane 1: Human HepG2 cell lysates, Lane 2: Human U87MG cell lysates, Lane 3: Human MG63 cell lysates, Lane 4: Human U2OS Cell Lysates, Primary: Anti-Osteopontin (orb500935) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 60 kDa, Observed band size: 60 kDa.

Paraformaldehyde-fixed, paraffin embedded (Rat lung), 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 (osteopontin) Polyclonal Antibody, Unconjugated (orb500935) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Lane 1: HepG2 (Human) Cell Lysate at 30 ug, Lane 2: A549 (Human) Cell Lysate at 30 ug, Lane 3: U251 (Human) Cell Lysate at 30 ug, Primary: Anti-Osteopontin (orb500935) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 60-65 kD, Observed band size: 60 kD.