Neurofilament 68 antibody | knockout validation | Sigma-Aldrich N5139

This is a knockout-validated antibody summary, based on the publication "Prion-like transmission of α-synuclein pathology in the context of an NFL null background", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Mouse monoclonal IgG1

Company: Sigma-Aldrich

Antibody: Neurofilament 68

Catalog number: N5139

Summary: Mouse monoclonal IgG1 against pig spinal cord. Reacts with rat, pig, chicken, and human. Suitable for western blot, immunohistochemistry (paraffin) and microarray.

Validation Method

Western blot

Sample

Spinal cord from wild-type and NFL null mice. Tissues were homogenized in 2 % sodium dodecyl sulfate (SDS)/50 mM Tris, pH 7.5 using a pestle, followed by probe sonication.

Blocking agent

5 % dry milk/Tris buffered saline (TBS).

Primary incubation

Diluted in 5 % dry milk/TBS overnight at 4℃.

Secondary incubation

Horseradish peroxidase conjugated goat anti-mouse secondary antibodies for 1 hour.

Detection

Western Lightning-Plus ECL reagents (PerkinElmer) and imaged on a GeneGnome XRQ system using GeneTools software (Syngene).

Figure

Western blot analysis of total protein lysates from spinal cord of wild-type (WT), M83, NFL null and M83/NFL null mice. Antibody NR4 shows the presence/absence of NFL. GAPDH and β-actin are shown as loading controls. Please see Figure 1b in the article [1].

Clone note

The same clone (NR4) is sold as Santa Cruz Biotechnology sc-58560; Biorbyt orb388518; Boster MA1070; Sigma-Aldrich N5139.

References
  1. Rutherford N, Brooks M, Riffe C, Gorion K, Howard J, Dhillon J, et al. Prion-like transmission of α-synuclein pathology in the context of an NFL null background. Neurosci Lett. 2017;661:114-120 pubmed publisher