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).

Guinea pig polyclonal IgG

Company: Progen

Antibody: NEPHRIN

Catalog number: GP-N2

Summary: Guinea pig polyclonal IgG against mouse myeloma cell line NS0-derived recombinant human Kirrel3/NEPH2 Tyr33-Ala535. Reacts with human and mouse. Suitable for western blot and immunohistochemistry (frozen and paraffin). For the latter, antigen-retrieval is recommended.

Western blot | Immunohistochemistry

Brain lysates from WT and Nephs1-KO ice. WB: tissues were glass-glass-homogenized in lysis buffer containing 20mM CHAPS and 1% Triton X-100. IHC: kidneys were frozen in OCT compound and sectioned at 5μm (Leica Kryostat, Wetzlar, Germany). The sections were fixed with 4% paraformaldehyde.

IHC: PBS containing 5% BSA + 5% Normal Donkey Serum (Jackson Immuno Research, Suffolk, UK).

IHC: for 45 min.

WB: HRP-coupled secondary antibodies.

IHC: fluorophore-conjugated secondary antibodies (Life Technologies, Darmstadt, Germany) for 30 min.

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

IHC: Zeiss 170 fluorescence microscope equipped with a 20x and 63x water immersion objective 171 (Zeiss, Oberkochen, Germany).

Please see Supplemental Figure 1i-k in the article [1].