Nephrin Antibody [Unconjugated]

Western blot shows lysates of human kidney tissue under reducing and non-reducing conditions. PVDF membrane was probed with 1 µg/mL Sheep Anti-Human Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4269) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band for Nephrin was detected at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Nephrin was detected in immersion fixed paraffin-embedded sections of human kidney using 1.7 µg/mL Sheep Anti-Human Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4269) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to podocytes in glomeruli. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western lane view shows lysates of human kidney, loaded at 0.2 mg/mL. A specific band was detected for Nephrin at approximately 185 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4269) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Nephrin Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Nephrin
  Gln23-Thr1029
  Accession # O60500
• Specificity: Detects human Nephrin in direct ELISAs and Western blots. In direct ELISAs, less than 20% cross-reactivity with recombinant mouse Nephrin is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: NPHS1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 1-15 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Nephrin Antibody [Unconjugated]

• CNF
• Nephrin
• nephrosis 1, congenital, Finnish type (nephrin)
• NPHNCNF
• NPHS1
• Renal glomerulus-specific cell adhesion receptor

Background

Nephrin, also known as renal glomerulus-specific cell adhesion receptor and nephrosis 1 (NPHS1) is a 185-200 kDa type I transmembrane protein belonging to the immunoglobulin (Ig) superfamily. It is expressed on podocytes and is an essential component of the interpodocyte-spanning slit diaphragm complex. Nephrin forms cis-hetero-oligomeric complexes with Neph1, followed by trans-homophilic interaction with Nephrin on opposing cells. Mutations in the Nephrin gene is the pathogenic cause of congenital nephrotic syndrome. Mature human Nephrin contains a 1033 aa extracellular region and a 165 aa cytoplasmic tail. One potential soluble splice form is known where aa’s 1056-1095 are deleted, eliminating the transmembrane region. Over aa 23-1029, human Nephrin shares 84% and 89% aa sequence identity with mouse and canine Nephrin, respectively.

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 Nephrin Antibody (AF4269)(21)

Publications using AF4269

• G Wang, B Heijs, S Kostidis, RGJ Rietjens, M Koning, L Yuan, GL Tiemeier, A Mahfouz, SJ Dumas, M Giera, J Kers, SM Chuva de S, CW van den Be, BM van den Be, TJ Rabelink Spatial dynamic metabolomics identifies metabolic cell fate trajectories in human kidney differentiation Cell Stem Cell, 2022;29(11):1580-1593.e7. 2022 [PMID: 36332571] (IHC, Human)
• AD Westerling, EM Fast, TW Soare, S Venkatacha, M DeRan, AB Fanelli, S Kyrychenko, H Hoang, GM Corriea, W Zhang, M Yu, M Daniels, G Malojcic, XR Pan-Zhou, MW Ledeboer, JC Harmange, M Emani, TT Tibbitts, JF Reilly, P Mundel Transplanted organoids empower human preclinical assessment of drug candidate for the clinic Science Advances, 2022;8(27):eabj5633. 2022 [PMID: 35857479] (Flow Cytometry, Human)
• N Won, J Castillo-P, X Tan, J Ford, D Heath, LI Mazilescu, M Selzner, IM Rogers Ex Vivo Perfusion Using a Mathematical Modeled, Controlled Gas Exchange Self-Contained Bioreactor Can Maintain a Mouse Kidney for Seven Days Cells, 2022;11(11):. 2022 [PMID: 35681517] (IHC, Mouse)
• J Jansen, KC Reimer, JS Nagai, FS Varghese, GJ Overheul, M de Beer, R Roverts, D Daviran, LAS Fermin, B Willemsen, M Beukenboom, S Djudjaj, S von Stillf, LE van Eijk, M Mastik, M Bulthuis, WD Dunnen, H van Goor, JL Hillebrand, SH Triana, T Alexandrov, MC Timm, BT van den Be, M van den Br, Q Nlandu, J Heijnert, EMJ Bindels, RM Hoogenboez, F Mooren, C Kuppe, P Miesen, K Grünberg, T Ijzermans, EJ Steenberge, J Czogalla, MF Schreuder, N Sommerdijk, A Akiva, P Boor, VG Puelles, J Floege, TB Huber, COVID Moon, RP van Rij, IG Costa, RK Schneider, B Smeets, R Kramann SARS-CoV-2 infects the human kidney and drives fibrosis in kidney organoids Cell Stem Cell, 2021;29(2):217-231.e8. 2021 [PMID: 35032430] (IHC, Human)
• L Miesen, P Bándi, B Willemsen, F Mooren, T Strieder, E Boldrini, V Drenic, J Eymael, R Wetzels, J Lotz, N Weiss, E Steenberge, TH van Kuppev, M van Erp, J van der La, N Endlich, MJ Moeller, JFM Wetzels, J Jansen, B Smeets Parietal epithelial cells maintain the epithelial cell continuum forming Bowman's space in focal segmental glomerulosclerosis Disease Models & Mechanisms, 2021;0(0):. 2021 [PMID: 34927672] (IHC, Rat)
• R Ungricht, L Guibbal, MC Lasbennes, V Orsini, M Beibel, A Waldt, R Cuttat, W Carbone, A Basler, G Roma, F Nigsch, JS Tchorz, D Hoepfner, PS Hoppe Genome-wide screening in human kidney organoids identifies developmental and disease-related aspects of nephrogenesis Cell Stem Cell, 2021;0(0):. 2021 [PMID: 34847364] (IHC, Human)
• KS Lee, J Lee, P Lee, BC Jeon, MY Song, S Kwak, J Lee, JS Kim, DJ Kim, JH Kim, VL Tesh, MS Lee, SK Park Inhibition of O-GlcNAcylation protects from Shiga toxin-mediated cell injury and lethality in host Embo Molecular Medicine, 2021;0(0):e14678. 2021 [PMID: 34842355] (IHC, Human)
• S Ueda, T Tominaga, A Ochi, A Sakurai, K Nishimura, E Shibata, S Wakino, M Tamaki, K Nagai TGF-beta1 is involved in senescence-related pathways in glomerular endothelial cells via p16 translocation and p21 induction Scientific Reports, 2021;11(1):21643. 2021 [PMID: 34737348] (IHC, Human)
• JW Kim, HW Kim, SA Nam, JY Lee, HJ Cho, TM Kim, YK Kim Human kidney organoids reveal the role of glutathione in Fabry disease Experimental & Molecular Medicine, 2021;0(0):. 2021 [PMID: 34654880] (IHC, Human)
• N Mangold, J Pippin, D Unnersjoe-, S Koehler, S Shankland, S Brähler, B Schermer, T Benzing, PT Brinkkoett, H Hagmann The Atypical Cyclin-Dependent Kinase 5 (Cdk5) Guards Podocytes from Apoptosis in Glomerular Disease While Being Dispensable for Podocyte Development Cells, 2021;10(9):. 2021 [PMID: 34572114] (IHC, Mouse)
• K Uchimura, H Wu, Y Yoshimura, BD Humphreys Human Pluripotent Stem Cell-Derived Kidney Organoids with Improved Collecting Duct Maturation and Injury Modeling Cell Reports, 2020;33(11):108514. 2020 [PMID: 33326782] (IHC, Human)
• J Usui, M Yaguchi, S Yamazaki, M Takahashi-, T Kawamura, S Kaneko, SV Seshan, P Ronco, K Yamagata Transcription factor 21 expression in injured podocytes of glomerular diseases Sci Rep, 2020;10(1):11516. 2020 [PMID: 32661376] (IHC, Human)
• Monteil V, Kwon H, Prado P, Hagelkruys A, Wimmer R, Stahl M, Leopoldi A, Garreta E, Hurtado Del Pozo C, Prosper F, Romero J, Wirnsberger G, Zhang H, Slutsky A, Conder R, Montserrat N, Mirazimi A, Penninger J Inhibition of SARS-CoV-2 Infections in Engineered Human Tissues Using Clinical-Grade Soluble Human ACE2. Cell, 0;181(4):905-913.e7. 0 [PMID: 32333836] (IHC, Human)
• A Subramania, EH Sidhom, M Emani, K Vernon, N Sahakian, Y Zhou, M Kost-Alimo, M Slyper, J Waldman, D Dionne, LT Nguyen, A Weins, JL Marshall, O Rosenblatt, A Regev, A Greka Single cell census of human kidney organoids shows reproducibility and diminished off-target cells after transplantation Nat Commun, 2019;10(1):5462. 2019 [PMID: 31784515] (ICC, Mouse)
• JL Harder, R Menon, EA Otto, J Zhou, S Eddy, NL Wys, C O'Connor, J Luo, V Nair, C Cebrian, JR Spence, M Bitzer, OG Troyanskay, JB Hodgin, RC Wiggins, BS Freedman, M Kretzler, , Organoid single cell profiling identifies a transcriptional signature of glomerular disease JCI Insight, 2019;4(1):. 2019 [PMID: 30626756] (IHC-P, Human)
• B Phipson, PX Er, AN Combes, TA Forbes, SE Howden, L Zappia, HJ Yen, KT Lawlor, LJ Hale, J Sun, E Wolvetang, M Takasato, A Oshlack, MH Little Evaluation of variability in human kidney organoids Nat. Methods, 2019;16(1):79-87. 2019 [PMID: 30573816] (IHC, Mouse)
• LJ Hale, SE Howden, B Phipson, A Lonsdale, PX Er, I Ghobrial, S Hosawi, S Wilson, KT Lawlor, S Khan, A Oshlack, C Quinlan, R Lennon, MH Little 3D organoid-derived human glomeruli for personalised podocyte disease modelling and drug screening Nat Commun, 2018;9(1):5167. 2018 [PMID: 30514835] (Human)
• H Wu, K Uchimura, EL Donnelly, Y Kirita, SA Morris, BD Humphreys Comparative Analysis and Refinement of Human PSC-Derived Kidney Organoid Differentiation with Single-Cell Transcriptomics Cell Stem Cell, 2018;0(0):. 2018 [PMID: 30449713] (IHC-Fr, Human)
• Bajaj P, Rodrigues A, Steppan C, Engle S, Mathialagan S, Schroeter T Human Pluripotent Stem Cell-Derived Kidney Model for Nephrotoxicity Studies. Drug Metab Dispos, 0;46(11):1703-1711. 0 [PMID: 30171163] (ICC, Human)
• ELMO1 protects renal structure and ultrafiltration in kidney development and under diabetic conditions Sci Rep, 2016;6(0):37172. 2016 [PMID: 27849017] (IHC-P, Human)
• Takasato M, Er P, Chiu H, Maier B, Baillie G, Ferguson C, Parton R, Wolvetang E, Roost M, Chuva de Sousa Lopes S, Little M Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis. Nature, 2015;526(7574):564-8. 2015 [PMID: 26444236] (IHC, Human)

Bioinformatics

• Gene Symbol: NPHS1
• Uniprot:
  • O60500()