Synthetic peptide within Human NPHS2 aa 334-383 / 383.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC, IF-Tissue
Target Molecular Weight
Predicted band size: 42 kDa
Positive Control
Human kidney tissue lysate, mouse kidney tissue lysate, rat kidney tissue lysates, human kidney tissue, mouse kidney tissue, rat kidney tissue, 293T.
Conjugation
unconjugated
Clone Number
JB51-33
Product Features
Form
Liquid
Concentration
Lot Concentration Lookup
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
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.
Background References
1. Volker L A et al. Characterization of a short isoform of the kidney protein podocin in human kidney. BMC Nephrol 14:102-102 (2013).
2. Boute N et al. NPHS2, encoding the glomerular protein podocin, is mutated in autosomal recessive steroid-resistant nephrotic syndrome. Nat Genet 24:349-354 (2000).
Sequence Similarity
Belongs to the band 7/mec-2 family.
Tissue Specificity
Almost exclusively expressed in the podocytes of fetal and mature kidney glomeruli.
This data was developed using ET7107-34, the same antibody clone in a different buffer formulation. Western blot analysis of NPHS2 on human kidney tissue lysate with Rabbit anti-NPHS2 antibody (ET7107-34) 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: ET7107-34, 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
This data was developed using ET7107-34, the same antibody clone in a different buffer formulation. Western blot analysis of NPHS2 on different lysates with Rabbit anti-NPHS2 antibody (ET7107-34) 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: ET7107-34, 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
This data was developed using ET7107-34, the same antibody clone in a different buffer formulation. 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 (ET7107-34, 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.
This data was developed using ET7107-34, the same antibody clone in a different buffer formulation. 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 (ET7107-34, 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.
This data was developed using ET7107-34, the same antibody clone in a different buffer formulation. 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 (ET7107-34, 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.
This data was developed using ET7107-34, the same antibody clone in a different buffer formulation. Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling NPHS2 (ET7107-34) 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 (ET7107-34, 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.
This data was developed using ET7107-34, the same antibody clone in a different buffer formulation. Flow cytometric analysis of NPHS2 was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-34, 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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