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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human LRRK2 aa 950 to the C-terminus. The exact sequence is proprietary.
Database link: Q5S007
Well-characterized antibodies to efficiently detect and purify LRRK2 protein are a critical need in the Parkinson's Disease (PD) research community. To help accelerate LRRK2 research, The Michael J. Fox Foundation (MJFF), working with Epitomics, Inc., has generated unique and high quality LRRK2 rabbit monoclonal antibodies to be widely available for PD research community.
LRRK2 (Leucine-rich repeat kinase 2, dardarin) is a protein kinase belonging to the ROCO family, which is defined by the presence of a ROC (Ras/GTPase of complex proteins) domain and COR (C-terminal of Roc) region. LRRK2 exhibits kinase activity whereby it can undergo autophosphorylation and can phosphorylate generic substrates. In addition, the GTPase domain of LRRK2 can mediate GDP (guanosine-5′-diphosphate)/GTP (guanosine-5′-triphosphate) binding as well as GTP hydrolysis.
LRRK2 is mutated in a significant number of Parkinson's disease (PD) patients. Mutations in this gene account for 4% of PD, and are observed in 1% of sporadic PD patients. Clinical symptoms of patients carrying PD-associated mutations of LRRK2 are indistinguishable from typical sporadic PD. The spectra of neuropathological features of PARK8 (type 8), the type corresponding to LRRK2, is broad and appears to encompass those associated with other familial PD cases such as PARK1 (alpha-synuclein) and PARK2 (Parkin). Patients with this gene mutation have typical relatively late onset Parkinsonism with features comparable with idiopathic PD; symptoms include asymmetric rest tremor, bradykinesia, rigidity, and a good response to 3,4-dihyroxy-l-phenylalanine (l-DOPA). The pathology of cases with LRRK2 mutations is pleomorphic.
This antibody clone (c68-7) has been re-introduced to the Epitomics catalog as characterization efforts have indicated that it performs very well at immunoprecipitating LRRK2 protein from cell lines and tissue lysate. This may facilitate investigators’ ability to more broadly examine alterations in LRRK2 protein levels and activity to interrogate changes in the biological samples.
For more characterization data and protocols using this LRRK2 Antibody, please refer to Davies, et al. 2013. Biochemical J 453(1):101-113 [PMID: 23560750]
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab181386 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration. PubMed: 26365310|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 286 kDa.
We recommend customers use 3% milk as the blocking agent for ab181386.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: LRRK2 knockout HAP1 cell lysate (20 µg)
Lane 3: MEF cell lysate (20 µg)
Lane 4: LRRK2 knockout MEF cell lysate (20 µg)
Lane 5: A549 cell lysate (20ug)
Lane 6: LRRK2 knockout A549 cell lysate (20 µg)
Lanes 1 - 6: Merged signal (red and green). Green - ab181386 observed at 238 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab181386 was shown to specifically react with wildtype HAP1, MEF and A549 cell lysates with additional cross-reactive bands. No bands were observed when LRRK2 knockout samples were used. Wild-type and LRRK2 knockout samples were subjected to SDS-PAGE. Membranes were blocked in 3% milk for 1 hour at room temperature. ab181386 and ab130007 (loading control to Vinculin) were diluted at 2.5 ug/mL and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Wild-type and LRRK2 knockout MEF and A549 cells were provide as a generous gift from Professor Dario Alessi, MRC Protein Phosphorylation and Ubiquitination Unit (University of Dundee).