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Nephrin Recombinant Rabbit Multiclonal Antibody [PSH07-70] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751182

Nephrin Recombinant Rabbit Multiclonal Antibody [PSH07-70] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Tissue

  • IHC-Fr

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Nephrin Recombinant Rabbit Multiclonal Antibody [PSH07-70] - BSA and Azide free

Antibody Type

Recombinant Rabbit multiclonal

Immunogen

Recombinant protein within human Nephrin aa 1-1,050.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Tissue, IHC-Fr

Molecular Weight

Predicted band size: 135 kDa

Positive Control

Mouse kidney tissue lysate, Rat kidney tissue lysate, human kidney tissue, mouse kidney tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

PSH07-70

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IHC-P

  • 1:3,000

  • IF-Tissue

  • 1:200-1:1,000

  • IHC-Fr

  • 1:500

Target

Function

Nephrin is a protein necessary for the proper functioning of the renal filtration barrier. The renal filtration barrier consists of fenestrated endothelial cells, the glomerular basement membrane, and the podocytes of epithelial cells. Nephrin is a transmembrane protein that is a structural component of the slit diaphragm. They are present on the tips of the podocytes as an intricate mesh and convey strong negative charges which repel protein from crossing into the Bowman's space. A defect in the gene for nephrin, NPHS1, is associated with congenital nephrotic syndrome of the Finnish type and causes massive amounts of protein to be leaked into the urine, or proteinuria. Nephrin is also required for cardiovascular development.

Background References

1. Watts AJB et al. Discovery of Autoantibodies Targeting Nephrin in Minimal Change Disease Supports a Novel Autoimmune Etiology. J Am Soc Nephrol. 2022 Jan

2. Hengel FE et al. Autoantibodies Targeting Nephrin in Podocytopathies. N Engl J Med. 2024 May

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: O60500 Human

SwissProt: Q9QZS7 Mouse

SwissProt: Q9R044 Rat

Synonyms

CNF antibody

Nephrin antibody

Nephrosis 1 congenital Finnish type antibody

Nephrosis 1, congenital, Finnish type (nephrin) antibody

NPHN antibody

NPHN_HUMAN antibody

NPHS 1 antibody

Nphs1 antibody

Renal glomerulus specific cell adhesion receptor antibody

Renal glomerulus-specific cell adhesion receptor antibody

Images

  • Immunofluorescence analysis of frozen mouse kidney tissue with Rabbit anti-Nephrin antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/500 dilution.<br /><br />The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.<br /><br />The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>, red) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of frozen mouse kidney tissue with Rabbit anti-Nephrin antibody (HA751182) at 1/500 dilution.

    The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.

    The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA751182, red) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Nephrin on different lysates with Rabbit anti-Nephrin antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/1,000 dilution.<br /><br />Lane 1: Mouse kidney tissue lysate<br />Lane 2: Mouse spleen tissue lysate (negative)<br />Lane 3: Rat kidney tissue lysate<br /><br />Lysates/proteins at 40 µg/Lane.<br /><br />Predicted band size: 135 kDa<br />Observed band size: 200 kDa<br /><br />Exposure time: 1 minute 30 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of Nephrin on different lysates with Rabbit anti-Nephrin antibody (HA751182) at 1/1,000 dilution.

    Lane 1: Mouse kidney tissue lysate
    Lane 2: Mouse spleen tissue lysate (negative)
    Lane 3: Rat kidney tissue lysate

    Lysates/proteins at 40 µg/Lane.

    Predicted band size: 135 kDa
    Observed band size: 200 kDa

    Exposure time: 1 minute 30 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751182) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Nephrin antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Nephrin antibody (HA751182) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751182) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Nephrin antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Nephrin antibody (HA751182) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751182) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Nephrin antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/3,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Nephrin antibody (HA751182) at 1/3,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751182) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Nephrin on different lysates with Rabbit anti-Nephrin antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/5,000 dilution.<br /><br />Lane 1: Mouse kidney tissue lysate<br />Lane 2: Mouse kidney tissue lysate treated with deglycosylation<br /><br />Lysates/proteins at 14.4 µg/Lane.<br /><br />Predicted band size: 135 kDa<br />Observed band size: 200/135 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751182" style="font-weight: bold;text-decoration: underline;">HA751182</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Nephrin on different lysates with Rabbit anti-Nephrin antibody (HA751182) at 1/5,000 dilution.

    Lane 1: Mouse kidney tissue lysate
    Lane 2: Mouse kidney tissue lysate treated with deglycosylation

    Lysates/proteins at 14.4 µg/Lane.

    Predicted band size: 135 kDa
    Observed band size: 200/135 kDa

    Exposure time: 3 minutes; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751182) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

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