NPHS2 Recombinant Rabbit Monoclonal Antibody [JB51-33]
cat.: HA750460
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, FC, IF-Tissue |
| Clonality: | Monoclonal |
| Clone number: | JB51-33 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 42 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human NPHS2 aa 334-383 / 383. |
| Positive control: | Human kidney tissue lysate, mouse kidney tissue lysate, rat kidney tissue lysates, human kidney tissue, mouse kidney tissue, rat kidney tissue, 293T. |
| Subcellular location: | Cell membrane. Endoplasmic reticulum. |
| Recommended Dilutions:
WB IHC-P FC IF-Tissue |
1:500-1:1,000 1:50-1:200 1:50-1:100 1:100 |
| Uniprot #: | SwissProt: Q9NP85 Human | Q91X05 Mouse | Q8K4G9 Rat |
| Alternative names: | Nephrosis 2, idiopathic, steroid resistant (podocin) nephrosis 2, idiopathic, steroid resistant NPHS2 NPHS2 gene PDCN PODO_HUMAN Podocin SRN1 |
Images
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Fig1:
Western blot analysis of NPHS2 on human kidney tissue lysate with Rabbit anti-NPHS2 antibody (HA750460) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; ECL: K1802 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA750460, 1/1,000 in 5% NFDM/TBST, 2 hours at room temperature Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 42 kDa Observed band size: 50 kDa |
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Fig2:
Western blot analysis of NPHS2 on different lysates with Rabbit anti-NPHS2 antibody (HA750460) at 1/1,000 dilution. Lane 1: Mouse kidney tissue lysate Lane 2: Rat kidney tissue lysate Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA750460, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 42 kDa Observed band size: 50 kDa |
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Fig3: 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 (HA750460, 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. |
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Fig4: 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 (HA750460, 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. |
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Fig5: 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 (HA750460, 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. |
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Fig6:
Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling NPHS2 (HA750460) and Vimentin (EM0401). 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 (HA750460, red) at 1/100 dilution and Vimentin (EM0401, green) at 1/400 dilution overnight at 4 ℃, washed with PBS. iFluor™ 594 conjugate-Goat anti-Rabbit IgG (HA1122) and iFluor™ 488 conjugate-Goat anti-Mouse IgG (HA1125) were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain. |
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Fig7: Flow cytometric analysis of NPHS2 was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (HA750460, 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). |
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