Podocin/NPHS2 Antibody (JB51-33)

Immunohistochemistry-Paraffin: Podocin/NPHS2 Antibody (JB51-33) [NBP2-75624] - Analysis of paraffin-embedded mouse kidney tissue using anti-Podocin/NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 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.

Flow Cytometry: Podocin/NPHS2 Antibody (JB51-33) [NBP2-75624] - Analysis of 293T cells with NPHS2 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.

Immunohistochemistry-Paraffin: Podocin/NPHS2 Antibody (JB51-33) [NBP2-75624] - Analysis of paraffin-embedded human kidney tissue using anti-Podocin/NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 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.

Immunohistochemistry-Paraffin: Podocin/NPHS2 Antibody (JB51-33) [NBP2-75624] - Rat kidney tissue using anti-NPHS2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 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.

Western Blot: Podocin/NPHS2 Antibody (JB51-33) [NBP2-75624] -Analysis of NPHS2 on rat kidney tissue lysates with Rabbit anti-NPHS2 antibody at 1/1,000 dilution.Lysates/proteins at 20 ug/Lane.Predicted band size: 42 kDaObserved band size: 50 kDaExposure time: 2 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 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.

Western Blot: Podocin/NPHS2 Antibody (JB51-33) [NBP2-75624] -Analysis of NPHS2 on different lysates with Rabbit anti-NPHS2 antibody at 1/500 dilution.Lane 1: Human kidney tissue lysateLane 2: Mouse kidney tissue lysateLysates/proteins at 20 ug/Lane.Predicted band size: 42 kDaObserved band size: 50 kDaExposure 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.

Immunohistochemistry-Paraffin: Podocin/NPHS2 Antibody (JB51-33) [NBP2-75624] -Immunofluorescence analysis of paraffin-embedded human kidney tissue labeling NPHS2 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 (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 (HA1125) were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IHC, mIF
• Clone: JB51-33
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Podocin/NPHS2 Antibody (JB51-33) Summary

• Description: Novus Biologicals Rabbit Podocin/NPHS2 Antibody (JB51-33) (NBP2-75624) is a recombinant monoclonal antibody validated for use in Multiplex Immunofluorescence, IHC, WB, Flow and ICC/IF. Anti-Podocin/NPHS2 Antibody: Cited in 4 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Synthetic peptide within Human Podocin/NPHS2 aa 334-383 / 383. (SwissProt: Q9NP85 Human; SwissProt: Q91X05 Mouse; SwissProt: Q8K4G9 Rat)
• Localization: Cell membrane. Endoplasmic reticulum.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: NPHS2
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:100
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Multiplex Immunofluorescence 1:1000
  • Western Blot 1:500-1:1000
• Theoretical MW: 42 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: TBS (pH7.4), 0.05% BSA, 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for Podocin/NPHS2 Antibody (JB51-33)

• Nephrosis 2
• nephrosis 2, idiopathic, steroid-resistant (podocin)
• NPHS2
• PDCN
• PDCNSRN1podocin
• Podocin
• SRN1

Background

NPHS2 encodes the glomerular protein podocin which plays a role in the regulation of glomerular permeability, and acts as a linker between the plasma membrane and the cytoskeleton. Defects in this gene are the cause of autosomal recessive steroid-resistant nephrotic syndrome (SRN). SRN is characterized by onset between three months and five years, resistance to steroid therapy and rapid progression to end-stage renal disease. An alternative splice variant has been described but its full length sequence has not been determined. [provided by RefSeq]

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Podocin/NPHS2 Antibody (NBP2-75624)(6)

Publications using NBP2-75624

• Radtke Aj, Chu Cj, Yaniv Z Et Al. IBEX: An open and extensible method for high content multiplex imaging of diverse tissues arXiv preprint 2021-01-01
• Tomita N, Hotta y, Naiki-Ito A et al. Protective effects of tadalafil on damaged podocytes in an adriamycin-induced nephrotic syndrome model Journal of Pharmacological Sciences 2022-03-01 [PMID: 35512855] (IHC-P, Rat)
• Pasupulati A, Nishad R, Mukhi D et al. Growth hormone induces TNF-alpha in podocytes and contributes to monocyte-to-macrophage differentiation: Implications in Diabetic kidney disease Research Square 2022-02-02 (WB, Human)
• Zhang Y, Yao H, Li C et al. Gandi Capsule Improved Podocyte Lipid Metabolism of Diabetic Nephropathy Mice through SIRT1/AMPK/HNF4A Pathway Oxidative medicine and cellular longevity 2022-04-18 [PMID: 35480869] (IHC-P, WB, Mouse)
• Ajay AK, Zhao L, Vig S et al. Deletion of STAT3 from Foxd1 cell population protects mice from kidney fibrosis by inhibiting pericytes trans-differentiation and migration Cell reports 2022-03-08 [PMID: 35263586] (IHC-P, Mouse)
• Radtke AJ, Chu CJ, Yaniv Z Et al. IBEX: an iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues Nat Protoc 2022-01-13 [PMID: 35022622] (Hydrolysis Assay, Human)
  
  Details:
  Citation using the Alexa Fluor 532 version of this antibody.

Bioinformatics

• Gene Symbol: NPHS2