Reports from the Literature: LRRK2 (N241A/34) as a Capture Antibody for ELISA

NeuroMab Clone N241A/34 is the Ideal Capture Antibody for Ultra-Sensitive LRRK2 ELISA Assays

In the first edition of our Antibodies in the Literature reports for 2025, we are pleased to update the utility of NeuroMab clone N241A/34, a mouse monoclonal antibody directed against the C-terminus of human LRRK2. Many of you are familiar with the use of clone N241A/34 for ICC (immunocytochemistry), IHC (immunohistochemistry), IP (immunoprecipitation) and WB (western blotting). Here we highlight recent reports in the literature that use NeuroMab clone N241A/34 as a capture antibody in ELISA assays for total LRRK2 and phospho-Ser935 LRRK2 with a limit of detection as low as 4 pg/mL.

Background of LRRK2

LRRK2 (Lucine Rich Repeat Kinase 2) is a protein coding gene that is associated with Parkinson’s Disease. LRRK2 is also known as Dardarin and PARK8, is a large multidomain protein kinase that includes leucine-rich repeats and a GTPase domain. The word “dardarin” is from the Basque word “dardara” that translates to English as “trembling.” Mutations in LRRK2 are commonly associated with late-onset Parkinson’s disease. LRRK2 is widely expressed and can be found in brain, kidney, lungs (1). LRRK2 is reported to be found in urine, cerebral spinal fluid, in lymphocytes and monocytes (2).

Structure and Function of LRRK2

The UniProt entry number for human LRRK2 is Q5S007 and the GeneCards symbol is LRRK2. Human LRRK2 has seven distinct domains: The protein is a serine/threonine-protein kinase which phosphorylates a broad range of proteins involved in multiple processes such as neuronal plasticity, innate immunity, autophagy, and vesicle trafficking. The mouse LRRK2 protein sequence is 87% homologous with the human amino acid sequence (see UniProt entry number Q5S006 for information about mouse LRRK2).

Figure 1. A schematic representation of the structure of LRRK2 with known pathogenic mutations designed by amino acid sequence numbers. There are seven domains in LRRK2 (ARM, Armadillo; ANK, Ankyrin repeat; LRR, leucine-rich repeat; ROC, Ras of complex proteins: GTPase; COR, C-terminal of ROC; WD40, WD-40 domain) and 2,527 amino acids.

LRRK2 Immunoassays Assays with 4 pg/mL Sensitivity

A Research Report published in the Journal of Parkinson’s Disease details using NeuroMab clone N241A/34 as the capture antibody for the development and validation of Quanterix SiMOA (“single molecule array”) ELISA assays for both total LRRK2 and phopho-Ser935 LRRK2. The limit of detection for the total LRRK2 assay was 12.1 pg/mL and 4.1 pg/mL for the phospho-Ser935 (3). The phosphorylated form of LRRK2, pSer935 LRRK2, has been proposed as a target modulation biomarker for LRRK2 inhibitors. In a 2018 study by the LRRK2 Detection in Human PBMC Consortium, researchers from Biogen, Denali, Merck, Pfizer, Sanofi and the Micheal J Fox Foundation developed a Quanterix SiMOA ELISA for total LRRK2 using NeuroMab clone N241A/34 at the capture antibody. This total LRRK2 assay had a limit of detection of 19 pg/mL. In parallel, they developed a Quanterix ELISA using NeuroMab clone N241A/34 as the capture antibody for the phospho-S935 LRRK2 assay, reported to have a limit of detection of 4.2 pg/mL (4).

Interestingly, the matrix for these studies was not serum, plasma or cerebral spinal fluid (csf) but rather a lysis buffer.  A buffer was used to lyse PBMC (peripheral blood mononuclear cells) isolated from whole blood. The lysis buffer containing the contents of lysed PBMC was then applied to the immunoassays for quantification of total LRRK2 and pSer935 LRRK2.

A 2024 study published in Molecular Neurodegeneration used NeuroMab clone N241A/34 as a capture antibody for developing a “single molecule array” immunoassay to measure LRRK2 in serum (5). A detailed protocol can be found here: SiMOA total LRRK2 Homebrew Assay.

NeuroMab clone N241A/34 is extensively cited in the literature.

Our NeuroMab clone N241A/34 is often cited in the literature – at least 150 times!  For a full list of citations, please click on this link and scroll down to the citations section.

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• Coating Buffers: Coating Buffers optimize antibody or antigen adsorption onto ELISA plates, stabilize tertiary structures, extend shelf-life, and minimize capture reagent usage.
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  • Sample Diluent Optimization Pack: Our optimization pack provides 4 x 100 mL bottles of Sample Diluent formulations (General Serum Diluent, Plasma Sample Diluent, Neptune Sample Diluent, and Protein-Free Sample Diluent) for an economical and fast method to screen for the best Sample Diluent for an assay.
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Citations

1. Westerlund M., Belin A.C., Anvret A., Bickford P., Olson L., Galter D. Developmental regulation of leucine-rich repeat kinase 1 and 2 expression in the brain and other rodent and human organs: Implications for Parkinson’s disease. Neuroscience. 2008;152(2):429–436.
2. Biskup S., Moore D.J., Rea A., Lorenz-Deperieux B., Coombes C.E., Dawson V.L., Dawson T.M., West A.B. Dynamic and redundant regulation of LRRK2 and LRRK1 expression. BMC Neurosci. 2007;8:102. 
3. Padmanabhan S, Lanz TA, Gorman D, Wolfe M, Joyce A, Cabrera C, Lawrence-Henderson R, Levers N, Joshi N, Ma TC, Liong C, Narayan S, Alcalay RN, Hutten SJ, Baptista MAS, Merchant K. An Assessment of LRRK2 Serine 935 Phosphorylation in Human Peripheral Blood Mononuclear Cells in Idiopathic Parkinson's Disease and G2019S LRRK2 Cohorts. J Parkinsons Dis. 2020;10(2):623-629.
4. PowerPoint Presentation
5. Yuan Y, Li H, Sreeram K, Malankhanova T, Boddu R, Strader S, Chang A, Bryant N, Yacoubian TA, Standaert DG, Erb M, Moore DJ, Sanders LH, Lutz MW, Velmeshev D, West AB. Single molecule array measures of LRRK2 kinase activity in serum link Parkinson's disease severity to peripheral inflammation. Mol Neurodegener. 2024 Jun 11;19(1):47.