This is a knockout-validated antibody summary, based on the publications "Detection of endogenous S1292 LRRK2 autophosphorylation in mouse tissue as a readout for kinase activity" [3] (western blot, see supplement figure 1a in the article), "A direct interaction between leucine-rich repeat kinase 2 and specific β-tubulin isoforms regulates tubulin acetylation" [1], "LRRK2 knockout mice have an intact dopaminergic system but display alterations in exploratory and motor co-ordination behaviors" [2] and "LRRK2 modifies α-syn pathology and spread in mouse models and human neurons" for western blot knockout validation (figure s7b) [4]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot of LRRK2 expression in WT and LRRK2 knock-out MEFs. α/β- tubulin was used as loading control. From [1].

Rabbit monoclonal

Company: Epitomics

Antibody: LRRK2

Catalog number: MJFF2, now Abcam ab133474

Summary: Rabbit monoclonal against a recombinant fragment within Human LRRK2 aa 950 to the C-terminus. Reacts with mouse, rat, and human. Suitable for western blot, immunoprecipitation, immunohistochemistry (paraffin and free floating), immunocytochemistry/immunofluorescence.

Western blot, Immunohistochemistry

Mouse skin primary fibroblast cells (MEF) were derived from the dorsal skin of postnatal day 0 (P0) wild-type or Lrrk2 knock-out mouse pups (38). Cells were harvested 24 h post-transfection in buffer containing 50 mm Tris, pH 7.5, 100 mm NaCl, 1% Triton X-100, 1× complete protease inhibitor mixture (Roche), and 1× Halt phosphatase inhibitor mixture (Pierce).

5% (w/v) skimmed milk in PBS plus 0.1% (v/v) Tween 20 or with 20% (v/v) horse serum in PBS.

1:2,000 dilution.

HRP-conjugated anti-rabbit secondary antibody (Santa Cruz) at a 1:2,000 dilution.

SuperSignal West Pico Chemiluminescent Substrate (Pierce).

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Figure 2. Immunohistochemistry with MJFF2 c41-2 antibody showing WT and KO brain sections at the level of the striatum. Specific signal is seen in the WT compared to KO. Rabbit IgG was used as an isotype control. Boxes depict enlarged images to the right. Scale bar 50 microns. From [2].

Immunohistochemistry

Mice brain sections from LRRK2 KO, HET, and WT genotypes. Tissues were formalin fixed, and paraffin embedded.

Dako All-purpose blocking solution for 30 minutes.

1:4,000 dilution for 45 min at room temperature.

Secondary antibodies from the Envision+ System Labeled Polymer HRP (Dako).

DAB substrate (Dako).