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    products/primary-antibodies/lrrk2-antibody-mjff3-c69-6-ab133475.pdf

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Neuroscience Neurology process Neurodegenerative disease Parkinson's disease Synuclein
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)

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Western blot - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)
  • Western blot - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)
  • Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [MJFF3 (c69-6)] to LRRK2
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Human

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Overview

  • Product name

    Anti-LRRK2 antibody [MJFF3 (c69-6)]
    See all LRRK2 primary antibodies
  • Description

    Rabbit monoclonal [MJFF3 (c69-6)] to LRRK2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Recombinant fragment within Human LRRK2 aa 950 to the C-terminus. The exact sequence is proprietary.

  • Positive control

    • IHC-P: Human brain hippocampus tissue WB: Jurkat cells transfected with LRRK2
  • General notes

    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]

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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 antibody was developed with support from The Michael J. Fox Foundation.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituent: 99% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    MJFF3 (c69-6)
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Parkinson's disease
    • Synuclein
    • Neuroscience
    • Diseases

Associated products

  • Alternative Versions

    • Anti-LRRK2 antibody [MJFF3 (c69-6)] - BSA and Azide free (ab183216)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133475 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P
1/200 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Predicted molecular weight: 286 kDa.
Notes
IHC-P
1/200 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Predicted molecular weight: 286 kDa.

Target

  • Function

    Positively regulates autophagy through a calcium-dependent activation of the CaMKK/AMPK signaling pathway. The process involves activation of nicotinic acid adenine dinucleotide phosphate (NAADP) receptors, increase in lysosomal pH, and calcium release from lysosomes. Together with RAB29, plays a role in the retrograde trafficking pathway for recycling proteins, such as mannose 6 phosphate receptor (M6PR), between lysosomes and the Golgi apparatus in a retromer-dependent manner. Regulates neuronal process morphology in the intact central nervous system (CNS). Plays a role in synaptic vesicle trafficking. Phosphorylates PRDX3. Has GTPase activity. May play a role in the phosphorylation of proteins central to Parkinson disease.
  • Tissue specificity

    Expressed in the brain. Expressed in pyramidal neurons in all cortical laminae of the visual cortex, in neurons of the substantia nigra pars compacta and caudate putamen (at protein level). Expressed throughout the adult brain, but at a lower level than in heart and liver. Also expressed in placenta, lung, skeletal muscle, kidney and pancreas. In the brain, expressed in the cerebellum, cerebral cortex, medulla, spinal cord occipital pole, frontal lobe, temporal lobe and putamen. Expression is particularly high in brain dopaminoceptive areas.
  • Involvement in disease

    Parkinson disease 8
  • Sequence similarities

    Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
    Contains 12 LRR (leucine-rich) repeats.
    Contains 1 protein kinase domain.
    Contains 1 Roc domain.
    Contains 7 WD repeats.
  • Domain

    The seven-bladed WD repeat region is critical for synaptic vesicle trafficking and mediates interaction with multiple vesicle-associated presynaptic proteins.
    The Roc domain mediates homodimerization and regulates kinase activity.
  • Post-translational
    modifications

    Autophosphorylated.
  • Cellular localization

    Membrane. Cytoplasm. Perikaryon. Mitochondrion. Golgi apparatus. Cell projection, axon. Cell projection, dendrite. Endoplasmic reticulum. Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane. Endosome. Lysosome. Mitochondrion outer membrane. Mitochondrion inner membrane. Mitochondrion matrix. Predominantly associated with intracytoplasmic vesicular and membranous structures (By similarity). Localized in the cytoplasm and associated with cellular membrane structures. Predominantly associated with the mitochondrial outer membrane of the mitochondria. Colocalized with RAB29 along tubular structures emerging from Golgi apparatus. Localizes in intracytoplasmic punctate structures of neuronal perikarya and dendritic and axonal processes.
  • Target information above from: UniProt accession Q5S007 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 120892 Human
    • Entrez Gene: 66725 Mouse
    • Entrez Gene: 300160 Rat
    • Omim: 609007 Human
    • SwissProt: Q5S007 Human
    • SwissProt: Q5S006 Mouse
    • Unigene: 187636 Human
    • Unigene: 37558 Mouse
    • Alternative names

      • augmented in rheumatoid arthritis 17 antibody
      • AURA17 antibody
      • Dardarin antibody
      • Leucine rich repeat kinase 2 antibody
      • leucine rich repeat serine threonine protein kinase 2 antibody
      • Leucine-rich repeat serine/threonine-protein kinase 2 antibody
      • LRRK 2 antibody
      • LRRK2 antibody
      • LRRK2_HUMAN antibody
      • PARK 8 antibody
      • PARK8 antibody
      • RIPK7 antibody
      • ROCO 2 antibody
      • ROCO2 antibody
      see all

    Images

    • Western blot - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)
      Western blot - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)

      Lane 1: A549 whole cell lysate (20 µg)
      Lane 2: MEF whole cell lysate (20 µg)
      Lane 3: A549 (KO) whole cell lysate (20 µg)
      Lane 4: MEF (KO) whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab133475 observed at 286 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab133475 was shown to specifically react with Wild type A549 and MEF cells whilst no band was observed when knockout samples were used. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and knockout samples were subjected to SDS-PAGE. Ab133475 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

       

    • Western blot - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)
      Western blot - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)This image is courtesy of Zhuohua Zhang Lab (Sanford-Burnham Medical Research Institute)
      All lanes : Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475) at 1/1000 dilution

      Lane 1 : HEK293 cell lysate transfected with 3*Flag vector
      Lane 2 : HEK293 cell lysate transfected with 3*Flag wild type, full length LRRK2

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 286 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)This image is courtesy of the Douglas Walker Lab

      ab133475, at 1/200 dilution, staining LRRK2 in paraffin embedded Human hippocampus tissue using immunohistochemical analysis.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)
      Anti-LRRK2 antibody [MJFF3 (c69-6)] (ab133475)

    Protocols

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab133475? Please let us know so that we can cite the reference in this datasheet.

    ab133475 has been referenced in 7 publications.

    • Penney J  et al. LRRK2 regulates retrograde synaptic compensation at the Drosophila neuromuscular junction. Nat Commun 7:12188 (2016). WB ; Mouse . PubMed: 27432119
    • Wandu WS  et al. Leucine-Rich Repeat Kinase 2 (Lrrk2) Deficiency Diminishes the Development of Experimental Autoimmune Uveitis (EAU) and the Adaptive Immune Response. PLoS One 10:e0128906 (2015). WB ; Mouse . PubMed: 26067490
    • Parisiadou L  et al. LRRK2 regulates synaptogenesis and dopamine receptor activation through modulation of PKA activity. Nat Neurosci 17:367-76 (2014). WB, ICC/IF ; Mouse . PubMed: 24464040
    • Davies P  et al. Comprehensive characterization and optimization of anti-LRRK2 (leucine-rich repeat kinase 2) monoclonal antibodies. Biochem J 453:101-13 (2013). WB, IP ; Mouse, Rat, Human . PubMed: 23560750
    • Häbig K  et al. LRRK2 guides the actin cytoskeleton at growth cones together with ARHGEF7 and Tropomyosin 4. Biochim Biophys Acta 1832:2352-2367 (2013). PubMed: 24075941
    • Wang X  et al. LRRK2 regulates mitochondrial dynamics and function through direct interaction with DLP1. Hum Mol Genet 21:1931-44 (2012). PubMed: 22228096
    • Thévenet J  et al. Regulation of LRRK2 expression points to a functional role in human monocyte maturation. PLoS One 6:e21519 (2011). WB ; Human . PubMed: 21738687

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