This is a knockout-validated antibody summary, based on the publication "The IkappaB kinase family phosphorylates the Parkinson's disease kinase LRRK2 at Ser935 and Ser910 during Toll-like receptor signaling", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Immunoblot of phospho-LRRK2 (Ser935) in HEK293 cell lysate containing overexpressed GFP-LRRK2 variants, LRRK2 wild type and knock-out mouse embryonic fibroblast (MEF) lysate and human lymphoblastoid cell lysate. Lymphoblast and MEF cells were treated plus or minus 1 µM LRRK2-IN1 (kinase inhibitor) for 2 h. Protein p38 served as loading control. From [1].

Rabbit monoclonal

Company: Epitomics, now part of Abcam

Antibody: Phospho LRRK2

Catalog number: 5099-1. The antibody is the same as Abcam ab133450

Summary: Rabbit monoclonal against a synthetic peptide corresponding to human LRRK2 (phospho S935). Reacts with mouse and human. Suitable for western blot and immunocytochemistry. Unsuitable for immunohistochemistry (paraffin).

Western blot

HEK293, LRRK2 wild type and knock-out mouse embryonic fibroblast (MEF) and human lymphoblastoid cells were lysed in buffer containing 50 mM Tris.HCL pH 7.5, 1 mM EGTA, 1 mM EDTA, 1 mM sodium orthovanadate, 10 mM β-glycerophosphate, 50 mM NAF, 5 mM sodium pyrophosphate, 0.27 M sucrose, 0.1% (v/v) 2-mercaptoethanol and 1% (v/v) Triton X-100.

5% (w/v) skimmed milk powder in TBST (Tris-buffered saline with Tween 20) buffer (50 mM Tris.HCl pH 7.5, 0.15 M NaCl and 0.1%(v/v) Tween 20)

100 ng/ml diluted in TBST with 5% (w/v) bovine serum albumin (BSA)

Anti-rabbit HRP (horseradish peroxidase) secondary antibodies (Sigma)

enhanced chemiluminescence reagent

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.