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<strong>Figure 1 Western Blot Validation in HeLa Cell Lysate with (A) the Absence and (B) the Presence of Blocking Peptide</strong><br>Loading: 15 μg of lysates per lane.Antibodies: Nephrin 7979 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Western Blot Validation in Human and Mouse Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: Nephrin 7979 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Immunohistochemistry Validation of Nephrin in Human Kidney Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Nephrin antibody (7979) 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.
<strong>Figure 4 Immunofluorescence Validation of Nephrin in Human Kidney Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed human kidney tissue labeling Nephrin with 7979 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 5 Immunofluorescence Validation of Nephrin in mouse podocyte clone 5 (MPC5) cells (Li et al, 2013) </strong><br> 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.
<strong>Figure 6 Induced Expression of Nephrin by curcumin treatment in the renal tissues of type 1 diabetic rats (Soetikno et al., 2013) </strong><br>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)
<strong>Figure 7 Immunofluorescence and Localization Validation of Nephrin in cultured rat podocytes (Piwkowska et al., 2012)</strong><br>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.
<strong>Figure 8 WB Validation of Nephrin in glomeruli of Zucker obese (ZO) and Zucker lean (ZL) rats (Piwkowska et al., 2013)</strong><br>The expression of nephrin detected by anti-nephrin antibodies did not change in ZO rats as compared to the control rats.
<strong>Figure 9 Immunohistochemistry Validation of Nephrin in mouse kidneys (Toyama et al., 2012) </strong><br>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.
<strong>Figure 10 Regulated Expression Validation of Nephrin in mouse podocyte cells cultured in normal glucose (NG) medium or high glucose (HG) medium (Huang et al., 2019) </strong><br>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.
Figure 1 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 7979 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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Nephrin Antibody

CATALOG NUMBER: 7979

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: Rat (93%).
Immunogen:
Anti-Nephrin antibody (7979) 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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
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.
Positive Control 1:
Cat. No. 1303 - Human Brain Tissue Lysate
Positive Control 2:
Cat. No. 1201 - HeLa Cell Lysate
Positive Control 3:
Cat. No. 1224 - Daudi Cell Lysate
Positive Control 4:
Cat. No. 1285 - C2C12 Cell Lysate
Predicted Molecular Weight:
Predicted: 125, 137 kDa
Observed: 125 kDa
Validation:

Induced Expression Validation (Figure 6): Nephrin expression detected by anit-Nephrin antibodies was up-regulated by curcumin treatment in type 1 diabetic rats.

Localization Validation (Figure 7): Nephrin detected by anti-Nephrin antibodies is colocalized with PKGIα at the tips of the cell processes in rat podocytes.

Regulated expression validation (Figure 10): Nephrin expression detected by anit-Nephrin antibodies was down-regulated by PEGF treatment (NGP or HGP), which was reversed by the addition of C3 transferase in mouse podocyte cells.

Isoforms:
Human Nephrin has 2 isoforms, including isoform 1 (1241aa, 135kD) and isoform 2 (1201aa, 131kD). Mouse Nephrin also has 2 isoforms, including isoform 1 (1256aa, 136kD) and isoform 2 (1242aa, 135kD). Rat Nephrin has 3 isoforms, including isoform 1 (1252aa, 136kD), isoform 2 (1239aa, 135kD) and isoform 3 (1166aa, 127kD). 7979 can detect human and mouse.
Purification:
Nephrin antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
Nephrin antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
Nephrin antibody can be stored at 4°C for three months and -20°C, stable for up to one year.
Ncbi Official Symbol:
NPHS1
Additional Names:
NPHN, NPHS1, Renal glomerulus-specific cell adhesion receptor3
Protein Accession Number:
NP_004637
Protein Gi Number:
4758822
Ncbi Gene Id Number:
4648
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
Nephrin is strongly expressed in renal glomeruli and is a member of the immunoglobulin family of cell adhesion molecules. Mutations in the Nephrin gene result in congenital nephrotic syndrome, an autosomal-recessive disorder characterized by massive proteinuria in utero and nephrosis at birth (1). Renal glomeruli allow normal kidneys to filter plasma so that it is very pure. Nephrin is expressed in the podocyte slit-diaphragm of the renal glomeruli in a manner that suggests that Nephrin molecules homodimerize in an anti-parallel fashion similar to cadherin interactions in adherens junctions. Thus, Nephrin may constitute the entire extracellular structure of the slit-diaphragm (2,3).
Background Reference 1:
Kestila et al. Mol. Cell 1998; 1:575-582.
Background Reference 2:
Tryggvason. J. Am. Soc. Nephrol. 1999; 10:2440-5.
Background Reference 3:
Tryggvason and Wartiovaara. Curr. Opin. Nephrol. Hypertens. 2001; 10:543-9.
Citation 1:
Li et al. Mesenchymal stem cells protect podocytes from apoptosis induced by high glucose via secretion of epithelial growth factor. Stem Cell Res Ther. 2013;4(5):103.PMID: 24004644
Citation 2:
Soetikno et al. Curcumin decreases renal triglyceride accumulation through AMPK-SREBP signaling pathway in streptozotocin-induced type 1 diabetic rats. J Nutr Biochem. 2013;24(5):796-802.PMID: 22898567
Citation 3:
Piwkowska et al. Insulin increases glomerular filtration barrier permeability through dimerization of protein kinase G type Iα subunits. Biochim Biophys Acta. 2013;1832(6):791-804. PMID: 23454089
Citation 4:
Toyama et al. Proteinuria in AMPD2-deficient mice. Genes Cells. 2012;17(1):28-38. PMID: 22212473
Citation 5:
Huang et al. Increased levels of serum pigment epithelium-derived factor aggravate proteinuria via induction of podocyte actin rearrangement. Int Urol Nephrol. 2019;51(2):359-367. PMID: 30536192
Citation 6:
Piwkowska et al. Hydrogen peroxide induces dimerization of protein kinase G type Iα subunits and increases albumin permeability in cultured rat podocytes. J Cell Physiol. 2012;227(3):1004-16. PMID: 21520075
Citation 7:
Rogacka et al. Metformin overcomes high glucose-induced insulin resistance of podocytes by pleiotropic effects on SIRT1 and AMPK. Biochim Biophys Acta Mol Basis Dis. 2018;1864(1):115-125. PMID: 29032153

FOR RESEARCH USE ONLY

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Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

7979

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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