Nephrin Rabbit Polyclonal Antibody

• Catalog Number: orb185637
• Category: Antibodies
• Description: Nephrin Rabbit Polyclonal Antibody
• Target: NPHS1
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Canine, Human, Mouse, Rat
• Predicted Reactivity: Rabbit
• 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 human Nephrin (451-550/1241aa)
• UniProt ID: O60500
• MW: 138 kDa
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/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: CNF; NPHN; Nephrosis 1 congenital Finnish type; NP
• Note: For research use only

Blank control: 293T. Primary Antibody (green line): Rabbit Anti-Nephrin antibody (orb185637), dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells wereincubated 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.

Blank control: RSC96 (blue), the cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice. Isotype Control Antibody: Rabbit IgG (orange), Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), dilution: 1:200 in 1X PBS containing 0.5% BSA, Primary Antibody, dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA (green).

Sample: Liver (Mouse) Lysate at 40 ug, Primary: Anti-Nephrin (orb185637) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 138 kD, Observed band size: 138 kD.

Sample: MDCK (Dog) Cell Lysate at 40 ug, Primary: Anti-Nephrin (orb185637) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 138 kD, Observed band size: 138 kD.

the cells (293T) were fixed with 2% paraformaldehyde (10 min). Isotype Control Antibody: Rabbit IgG (orange), Secondary Antibody: Goat anti-rabbit IgG-PE (white blue) dilution: 1:200 in 1X PBS containing 0.5% BSA, Primary Antibody Dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA (green).

Tissue/Cell: mouse kidney tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), 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-Nephrin Polyclonal Antibody, Unconjugated (orb185637) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: rat kidney tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), 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-Nephrin Polyclonal Antibody, Unconjugated (orb185637) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.