NPHS1 Antibody

• Catalog Number: orb1239658
• Category: Antibodies
• Description: NPHS1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, WB
• Predicted Reactivity: Rat
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: Anti-Nephrin antibody (orb1239658) was raised against a peptide corresponding to 15 amino acids near the center of human Nephrin. The immunogen is located within amino acids 1100-1150 of Nephrin.
• Concentration: 1 mg/mL
• Dilution range: WB: 1-2 μg/mL; IHC: 5 μg/mL; IF: 20 μg/mL.Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 125, 137 kDa Observed: 125 kDa
• Target: NPHS1
• UniProt ID: O60500
• NCBI: NP_004637
• Storage: Nephrin antibody can be stored at 4°C for three months and -20°C, stable for up to one year.
• Buffer/Preservatives: Nephrin antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: NPHN, NPHS1, Renal glomerulus-specific cell adhesi
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot Validation in HeLa Cell Lysate with (A) the Absence and (B) the Presence of Blocking Peptide. Loading: 15 µg of lysates per lane. Antibodies: Nephrin orb1239658 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Human and Mouse Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: Nephrin orb1239658 (2 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunohistochemistry Validation of Nephrin in Human Kidney Tissue. Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Nephrin antibody (orb1239658) at 5 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunofluorescence Validation of Nephrin in Human Kidney Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed human kidney tissue labeling Nephrin with orb1239658 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Immunofluorescence Validation of Nephrin in mouse podocyte clone 5 (MPC5) cells (Li et al, 2013). Double immunofluorescence analysis of podocytic membrane protein nephrin (red) and nuclei stained with DAPI (blue). The presence of high glucose (HG) and neutralizing antibody (NtAb) which blocked epithelial growth factor (EGF) decreased nephrin expression while mesenchymal stem cells-conditioned medium (MSCs-CM) and recombinant human EGF (rhEGF) prevented the effect.

Induced Expression of Nephrin by curcumin treatment in the renal tissues of type 1 diabetic rats (Soetikno et al., 2013). Nephrin expression detected by anti-nephrin antibodies in type 1 diabetic rats. Nephrin was down-regulated in the vehicle-treated diabetic rats as compared to the control nondiabetic rats. However, this decreasein nephrin protein expression was markedly increased by curcumin treatment (P<.05) to near-normal levels. (n=5 in each group).

Immunofluorescence and Localization Validation of Nephrin in cultured rat podocytes (Piwkowska et al., 2012). Immunofluorescence staining showed Nephrin expression (green) detected by anti-nephrin antibodies and PKGIalpha (red). The co-localization of two antibodies (yellow) in rat podocytes was observed particularly at the tips of the cell processes.

WB Validation of Nephrin in glomeruli of Zucker obese (ZO) and Zucker lean (ZL) rats (Piwkowska et al., 2013). The expression of nephrin detected by anti-nephrin antibodies did not change in ZO rats as compared to the control rats.

Immunohistochemistry Validation of Nephrin in mouse kidneys (Toyama et al., 2012). Protein analysis for nephrin by immunohistochemistry with anti-nephrin antibodies in kidneys of wild-type or AMPD2-deficient mice at 2, 12 or 24 weeks of age. No difference between wild-type andAMPD2-deficient mice at any age was observed.

Regulated Expression Validation of Nephrin in mouse podocyte cells cultured in normal glucose (NG) medium or high glucose (HG) medium (Huang et al., 2019). Western Blot analysis was used to access the protein expression level of nephrin with anti-nephrin antibodies. Nephrin expression was down-regulated by PEGF treatment (NGP or HGP), which was reversed by the addition of C3 transferase.