NEPH3 antibody | knockout validation | R&D AF2930
DOI
//dx.doi.org/10.13070/ko.en.7.2197
Date
2017-03-16

This is a knockout-validated antibody summary, based on the publication "A flexible, multilayered protein scaffold maintains the slit in between glomerular podocytes", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Goat polyclonal IgG

Company: R&D

Antibody: NEPH3

Catalog number: AF2930

Summary: Goat polyclonal IgG against mouse myeloma cell line NS0-derived recombinant mouse Kirrel2/NEPH3

Ser19-Asp501. Reacts with mouse by western blot.

Validation Method

Western blot

Sample

Brain lysates from WT and Neph3-KO ice. Tissues were glass-glass-homogenized in RIPA buffer containing: 50mM Tris/HCl pH 7,5, 1 mM EGTA, 1 mM EDTA, 1% (w/v) Triton X-100, 0,1 % (w/v) SDS, 50 mM NaF, 150 mM NaCl, 0,5% (w/v) Na-Deoxycholate, 0,1 %(v/v) 2-mercaptoethanol, 1mM Na-Orthovanadate, Roche Ultra complete proteinase inhibitor cocktail and Roche Phospho-STOPP as indicated by the manufacturer (Roche, Mannheim, Germany) and ddH2O ad final volume; 15μl lysis buffer per 1 mg of tissue were used).

Secondary incubation

HRP-coupled secondary antibodies.

Detection

ECL in combination with a conventional x-ray system (films: Fuji, Tokio, Japan; developer: AGFA, Mortsel, Belgium).

Figure

Please see Supplemental Figure 2d in the article [1].

References
  1. Grahammer F, Wigge C, Schell C, Kretz O, Patrakka J, Schneider S, et al. A flexible, multilayered protein scaffold maintains the slit in between glomerular podocytes. JCI Insight. 2016;1: pubmed