Anti-LRRK2 antibody [MJFF2 (c41-2)] is a rabbit recombinant monoclonal antibody that is used to detect LRRK2 in IP, Western blot. Suitable for Human, Mouse samples.
- Cited in over 120 publications
- Specificity confirmed with LRRK2 knockout cell line validation
- LRRK2 MJFF2 (c41-2) was developed with support from the Michael J. Fox Foundation for Parkinson’s Research
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IP | WB | ICC/IF | |
---|---|---|---|
Human | Tested | Tested | Not recommended |
Mouse | Tested | Tested | Not recommended |
Rat | Predicted | Predicted | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes (2-5 μg) |
Species Human | Dilution info 1/60 | Notes (2-5 μg) |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/10000 - 1/50000 | Notes - |
Species Human | Dilution info 1/10000 - 1/50000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info - | Notes - |
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The protein expressed by the gene LRRK2 is a serine/threonine-protein kinase that phosphorylates a wide range of proteins involved in neuronal plasticity, autophagy, and vesicle trafficking. It serves as a key regulator of RAB GTPases by affecting the GTP/GDP exchange and interaction partners of RABs through phosphorylation, targeting proteins like RAB3A, RAB3B, and others. LRRK2 manages the GDP/GTP exchange for RAB8A by phosphorylating 'Thr-72', inhibiting RAB8A's interaction with GDI1/GDI2, and influencing primary ciliogenesis to enhance SHH signaling in the brain. It works with RAB29 in retromer-dependent recycling of proteins between lysosomes and the Golgi. The protein is involved in shaping neuronal morphology in the CNS, synaptic vesicle trafficking, and recruiting SEC16A to assist in ER to Golgi transport. It enhances autophagy via the CaMKK/AMPK pathway and relates to nicotinic acid adenine dinucleotide phosphate receptors and lysosomal activity. LRRK2 phosphorylates PRDX3 and APP, influencing neuron apoptosis. Independently, it inhibits MAPT degradation, fostering MAPT oligomerization. Additionally, LRRK2 possesses GTPase activity that regulates its kinase activity. This supplementary information is collated from multiple sources and compiled automatically.
PARK8, LRRK2, Leucine-rich repeat serine/threonine-protein kinase 2, Dardarin
Anti-LRRK2 antibody [MJFF2 (c41-2)] is a rabbit recombinant monoclonal antibody that is used to detect LRRK2 in IP, Western blot. Suitable for Human, Mouse samples.
- Cited in over 120 publications
- Specificity confirmed with LRRK2 knockout cell line validation
- LRRK2 MJFF2 (c41-2) was developed with support from the Michael J. Fox Foundation for Parkinson’s Research
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Product Specifications
Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in IP, WB in human, mouse samples.
Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) specifically detects LRRK2 (UniProt ID: Q5S007; Molecular weight: 286kDa) and is sold in 100 µL and 1 mL selling sizes.
Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) was developed by Abcam in partnership with the Michael J. Fox Foundation for Parkinson's Research.
Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) has been confirmed by testing in knockout samples.
Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) has been cited over 120 times in peer reviewed journals and is trusted by the scientific community.
Related Products
Conjugation-ready, carrier free format available for antibody clone MJFF2 (c41-2) - Anti-LRRK2 antibody [MJFF2 (c41-2)] - BSA and Azide free ab172378.
Antibody clone MJFF2 (c41-2) is also available pre-conjugated to a variety of labels for your convenience - HRP (HRP Anti-LRRK2 antibody [MJFF2 (c41-2)] ab195024).
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 Abcam, 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.
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]
Abcam recommended secondaries - Goat Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Or search our wide range of secondary antibodies for use with your experiment.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Collaborations
This antibody was developed with support from The Michael J. Fox Foundation.
The protein LRRK2 also known as leucine-rich repeat kinase 2 or dardarin is an enzyme with a molecular weight of approximately 286 kDa. It functions as a kinase meaning it adds phosphate groups to other proteins which affects their activity. LRRK2 is expressed in various tissues but it is highly abundant in the brain especially in regions such as the striatum and cortex. It has a significant role in cellular signaling processes due to its phosphorylation activity.
LRRK2 interacts with cellular mechanisms by regulating cytoskeletal dynamics autophagy and vesicle trafficking. It is a part of a larger complex that includes other proteins involved in these processes. The kinase activity of LRRK2 plays an essential part in maintaining neuronal health and function. It influences the process of autophagy which is a way cells clean themselves by removing damaged components and recycling them.
The action of LRRK2 is central to the mitogen-activated protein kinase (MAPK) and the mammalian target of rapamycin (mTOR) pathways. In these pathways LRRK2 interacts with other proteins such as mTOR and RPS6KB1. It modulates cellular processes like growth proliferation and response to stressors. Its kinase activity affects the phosphorylation state of targets within the pathways hence influencing biological outcomes like survival and apoptosis.
LRRK2 mutations have a significant connection to Parkinson's disease and Crohn's disease. In Parkinson's disease mutated LRRK2 leads to abnormal protein aggregation linking to proteins such as alpha-synuclein. For Crohn's disease LRRK2 influences the immune response and intestinal inflammation. These connections highlight LRRK2's role in the pathogenesis and contribute to understanding these complex disorders.
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Lanes 1 - 4: Merged signal (red and green). Green - ab133474 observed at 286 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab133474 was shown to recognize LRRK2 in wild type A549 and MEF cells along with additional cross reative bands. Whilst signal was not seen in LRRK2 knockout cells. Wild-type and LRRK2 knockout samples were subjected to SDS-PAGE. ab133474 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 10000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed 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).
All lanes: Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
Predicted band size: 286 kDa
Immunoprecipitation to verify the interaction of LRRK2 and ArfGAP1 in vivo. LRRK2 interacts with ArfGAP1 in brain extracts derived from wild-type mice following immunoprecipitation with ab133474, a LRRK2-specific monoclonal antibody (MJFF-2/c41-2), whereas ArfGAP1 is not immunoprecipitated in extracts derived from LRRK2 knockout mice
Protein extracts were prepared from the cerebral cortex of adult wild-type and LRRK2 knockout mice (with targeted deletion of exon 41 of the LRRK2 gene) by homogenization in TNE buffer (10 mM Tris-HCL pH 7.4, 150 mM NaCl, 5 mM EDTA, 0.5% NP-40, 1× phosphatase inhibitor cocktail 1 and 2, 1× Complete Mini protease inhibitor cocktail). Protein concentration was determined by BCA assay. Brain extracts (10 mg protein) were combined with 50 μl Protein G-Dynabeads pre-incubated with rabbit anti-LRRK2 (5 μg; MJFF2/c41-2; Abcam, Inc.), rabbit anti-ArfGAP1 (3 μg) or rabbit IgG (3 μg) antibodies followed by overnight incubation at 4°C. Dynabead complexes were sequentially washed twice with TNE buffer and twice with TBS buffer (10 mM Tris-HCL pH 7.4, 150 mM NaCl). Immunoprecipitates were eluted by heating at 70°C for 10 min, resolved by SDS-PAGE and subjected to Western blot analysis.
All lanes: Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
Predicted band size: 286 kDa
LRRK2 was immunoprecipitated from 0.35 mg A549 (Human lung carcinoma epithelial cell) whole cell lysate 10 μg with 133474 at 1/60 dilution (2μg). VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: A549 (Human lung carcinoma epithelial cell) whole cell lysate 10 μg
Lane 2: ab133474 IP in A549 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab133474 in A549 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Fresh lysate should be used to minimize protein degradation.
All lanes: Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
Predicted band size: 286 kDa
Observed band size: 286 kDa
All lanes: Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) at 1/10000 dilution
Lane 1: HEK293 cell lysate transfected with 3*Flag vector at 10 µg
Lane 2: HEK293 cell lysate transfected with 3*Flag full length wild type LRRK2 at 10 µg
All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 286 kDa
ab133474 was shown to react with LRRK2 in wild-type A549 cells in Western blot with loss of signal observed in a LRRK2 knockout cell line. Wild-type A549 and LRRK2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab133474 overnight at 4 °C at a 1/10000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) at 1/10000 dilution
Lane 1: Wild-type A549 lysate at 25 µg
Lane 2: LRRK2 knock-out A549 lysate at 25 µg
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