WB
Western blot analysis of NPHS2 on rat kidney tissue lysates with Rabbit anti-NPHS2 antibody at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of NPHS2 on different lysates with Rabbit anti-NPHS2 antibody at 1/500 dilution. Lane 1: Human kidney tissue lysate, Lane 2: Mouse kidney tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 4 minutes; 10% 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 at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.IF-P
Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling NPHS2 and Vimentin. 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 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies NPHS2 (red) at 1/100 dilution and Vimentin (green) at 1/400 dilution overnight at 4 ℃, washed with PBS. iFluor™ 594 conjugate-Goat anti-Rabbit IgG and iFluor™ 488 conjugate-Goat anti-Mouse IgG were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain.mIHC
Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-NPHS2 (Red), anti-AQP1 (Green), anti-Laminin beta 1 (Cyan) and anti-aSMA (Magenta) on kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit. The section was incubated in four rounds of staining: in the order of anti-NPHS2 (1/1,000 dilution), anti-AQP1 (1/5,000 dilution), anti-Laminin beta 1 (1/1,000 dilution) and anti-aSMA (1/10,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain.FC
Flow cytometric analysis of NPHS2 was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | NPHS2 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Plays a role in the regulation of glomerular permeability, acting probably as a linker between the plasma membrane and the cytoskeleton. Almost exclusively expressed in the podocytes of fetal and mature kidney glomeruli. |
| Immunogen | Synthetic peptide within Human NPHS2 aa 334-383 / 383. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCIF-PIHCWBmIHC |
| WB:1:500-1:1,000 IHC:1:50-1:200 FC:1:50-1:100 IF-P:1:100 mIHC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Nephrosis 2 idiopathic steroid resistant (podocin) antibody nephrosis 2 idiopathic steroid resistant antibody NPHS2 antibody NPHS2 gene antibody PDCN antibody PODO_HUMAN antibody Podocin antibody SRN1 antibody |
|---|---|
| Molecular Weight(MW) | 42 kDa(Observed band size: 50 kDa) |
| Cellular Localization | Cell membrane. Endoplasmic reticulum. |
WB
Western blot analysis of NPHS2 on rat kidney tissue lysates with Rabbit anti-NPHS2 antibody at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of NPHS2 on different lysates with Rabbit anti-NPHS2 antibody at 1/500 dilution. Lane 1: Human kidney tissue lysate, Lane 2: Mouse kidney tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 4 minutes; 10% 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 at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.
IF-P
Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling NPHS2 and Vimentin. 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 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies NPHS2 (red) at 1/100 dilution and Vimentin (green) at 1/400 dilution overnight at 4 ℃, washed with PBS. iFluor™ 594 conjugate-Goat anti-Rabbit IgG and iFluor™ 488 conjugate-Goat anti-Mouse IgG were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain.
mIHC
Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-NPHS2 (Red), anti-AQP1 (Green), anti-Laminin beta 1 (Cyan) and anti-aSMA (Magenta) on kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit. The section was incubated in four rounds of staining: in the order of anti-NPHS2 (1/1,000 dilution), anti-AQP1 (1/5,000 dilution), anti-Laminin beta 1 (1/1,000 dilution) and anti-aSMA (1/10,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain.
FC
Flow cytometric analysis of NPHS2 was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:500-1:1,000 IHC:1:50-1:200 FC:1:50-1:100 IF-P:1:100 mIHC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
