LRRK2 antibody | knockout validation | Epitomics 5097-1
DOI
//dx.doi.org/10.13070/ko.en.6.1714
Date
2016-10-07

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).

LRRK2  antibody | knockout validation | Epitomics 5097-1 figure 1
Figure 1. Immunoblot of LRRK2 in wild type and LRRK2 knock-out (KO) mouse embryonic fibroblast (MEF) lysate treated plus or minus 1 µM LRRK2-IN1 (kinase inhibitor) for 2 h. Protein p38 served as loading control. From [1].
Antibody information

Rabbit monoclonal

Company: Epitomics, Now part of Abcam.

Antibody: LRRK2

Catalog number: 5097-1, the antibody is the same as Abcam ab133518.

Summary: Rabbit monoclonal against a synthetic peptide corresponding to human LRRK2 (aa 100-500). Reacts with mouse and human.Suitable for western blot, immunocytochemistry, and immunohistochemistry (paraffin). Unsuitable for immunoprecipitation.

Validation Method

Western blot

Sample

LRRK2 wild type and knock-out mouse embryonic fibroblast (MEF) 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.

Blocking agent

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)

Primary incubation

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

Secondary incubation

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

Detection

enhanced chemiluminescence reagent

Disclaimer

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.

References
  1. Dzamko N, Inesta-Vaquera F, Zhang J, Xie C, Cai H, Arthur S, et al. The IkappaB kinase family phosphorylates the Parkinson's disease kinase LRRK2 at Ser935 and Ser910 during Toll-like receptor signaling. PLoS ONE. 2012;7:e39132 pubmed publisher