SPARC Rabbit Polyclonal Antibody

• Catalog Number: orb11405
• Category: Antibodies
• Description: SPARC Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human SPARC (101-200/303aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 33 kDa
• Target: SPARC
• UniProt ID: P09486
• RRID: AB_10751984
• 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: AA517111; Basement membrane protein 40; BM 40; BM4
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: U87MG. Primary Antibody (green line): Rabbit Anti-SPARC antibody (orb11405), Dilution: 2 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 0.5 ug/Test. Protocol, The cells were 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.

Formalin-fixed and paraffin embedded human ovarian tissue labeled with Anti-SPARC Polyclonal Antibody, Unconjugated (orb11405) at 1:200 followed by conjugation to the secondary antibody and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human brain glioma), 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 (SPARC) Polyclonal Antibody, Unconjugated (orb11405) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma), 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 (SPARC) Polyclonal Antibody, Unconjugated (orb11405) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human kidney), 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 (SPARC) Polyclonal Antibody, Unconjugated (orb11405) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human liver), 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 (SPARC) Polyclonal Antibody, Unconjugated (orb11405) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human rectal carcinoma), 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 (SPARC) Polyclonal Antibody, Unconjugated (orb11405) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue), 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 (SPARC) Polyclonal Antibody, Unconjugated (orb11405) at 1:400 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, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (SPARC) Polyclonal Antibody, Unconjugated (orb11405) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: A549 (Human) Cell Lysate at 30 ug, Primary: Anti-SPARC (orb11405) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 33 kD, Observed band size: 35 kD.

Sample: Brain (Mouse) Lysate at 40 ug, Primary: Anti-SPARC (orb11405) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 33 kD, Observed band size: 33 kD.

Tissue/Cell: Human Meningioma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-SPARC Polyclonal Antibody, Unconjugated (orb11405) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: mouse kidney tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-SPARC Polyclonal Antibody, Unconjugated (orb11405) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (orb868589) used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 ug/ml, blue) was used to stain the cell nuclei.

Tissue/Cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-SPARC Polyclonal Antibody, Unconjugated (orb11405) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (orb868589) used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 ug/ml, blue) was used to stain the cell nuclei.