Overview

  • Product name
    Anti-LRRK2 antibody [UDD3 30(12)]
    See all LRRK2 primary antibodies
  • Description
    Rabbit monoclonal [UDD3 30(12)] to LRRK2
  • Specificity
    This antibody does not give a positive signal in Human fetal brain. Please contact our Scientific Support team if you have any questions.
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WBmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human LRRK2 aa 100-500.

  • Positive control
    • GFP-LRRK2, GFP LRRK2 S910A, GFP LRRK2 S935A, LRRK2 WT MEF, LRRK2 WT MEF lysate from LRRK2-IN1 treated cells, Lymphoblastoid lysates and Lymphoblastoid lysate from LRRK2-IN1 treated cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    In recent years, a critical need in the Parkinson's Disease (PD) research community has been access to well-characterized antibodies that can be used to efficiently detect and purify Leucine-Rich Repeat Kinase 2 (LRRK2) protein. The Michael J. Fox Foundation (MJFF) has supported this effort by partnering with Dr. Dario Alessi (MRC Protein Phosphorylation Unit, University of Dundee) to help accelerate LRRK2 research. Dr. Alessi has characterized unique and high quality LRRK2 rabbit monoclonal antibodies, generated by Epitomics, to be made widely available for PD research community.

    LRRK2 (Leucine-rich repeat kinase 2, dardarin) is a multi-domain protein belonging to the ROCO family of proteins that contains a kinase and GTPase domain among its many protein interaction domains. 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. The most common mutation replaces glycine 2019 with a serine that results in increased LRRK2 kinase activity. This indicates that inhibitors of LRRK2 kinase activity might be of therapeutic benefit for the treatment of Parkinson’s disease and has stimulated much activity in this field of research.

    The Dundee-MJFF LRRK2[100-500] total antibody will be of great utility in further understanding LRRK2 as it relates to Parkinson’s disease. It should be noted this antibody is highly selective and sensitive and can readily be used to monitor LRRK2 protein levels in immunoblot analysis of 2-20 microgram amounts of whole cell extract. The LRRK2[100-500] total antibody recognizes both mouse and human endogenous LRRK2.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at 4°C prior to reconstitution. Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer
    pH: 7.40
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    UDD3 30(12)
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab133518 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
ICC/IF 1/200.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

 

WB 1/1000 - 1/10000. Predicted molecular weight: 286 kDa.
  • Application notes
    Is unsuitable for IP.
  • 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.
    • Information by UniProt
    • Database links
    • 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



    • Predicted band size : 286 kDa

      Lane 1: Wild-type A549 cell lysate (20 µg)
      Lane 2: LRRK2 knockout A549 cell lysate (20 µg) 
      Lane 3: Wild-type MEF cell lysate (20 µg)
      Lane 4: LRRK2 knockout MEF cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab133518 observed at 238 kDa. Red - loading control, ab130007, observed at 124 kDa.

      ab133518 was shown to specifically react with wild type A549 and MEF cell lines. No band was observed when knock out samples were used. Wild-type and LRRK2 knockout samples were subjected to SDS-PAGE. Ab133518 and ab130007 (loading control to Vinculin) were diluted at 1/500 and 1/10000 dilution respectively and incubated overnight at 4C. 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 generous a gift from Professor Dario Alessi, MRC Protein Phosphorylation and Ubiquitination Unit (University of Dundee).

    • Immunohistochemical staining of paraffin-embedded human kidney with purified ab133518 at a dilution of 1/100. A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • Immunofluorescent staining of SH-SY5Y cells fixed and permeablized with 4% PFA and 0.1% Triton X 100 using purified ab133518 at a dilution of 1/200. An Alexa Fluor® 488 goat anti-rabbit was used as the secondary (ab150077, 1/400) and the sample was stained with DAPI. The negative control is shown in bottom right hand panel - for the negative control, purified ab133518 was used at a dilution of 1/200 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500.

    • All lanes : Anti-LRRK2 antibody [UDD3 30(12)] (ab133518) at 1/1000 dilution (Unpurified)

      Lane 1 : GFP-LRRK2 lysate at 5 µg
      Lane 2 : GFP LRRK2 S910A lysate at 5 µg
      Lane 3 : GFP LRRK2 S935A lysate at 5 µg
      Lane 4 : LRRK2 WT MEF lysate at 20 µg
      Lane 5 : LRRK2 WT MEF lysate from LRRK2-IN1 treated cells at 20 µg
      Lane 6 : LRRK2 KO MEF lysate at 20 µg
      Lane 7 : LRRK2 KO MEF lysate from LRRK2-IN1 treated cells at 20 µg
      Lane 8 : Lymphoblastoid lysate at 30 µg
      Lane 9 : Lymphoblastoid lysate from LRRK2-IN1 treated cells at 30 µg

      Secondary
      Goat-anti-rabbit HRP at 1/2000 dilution

      Predicted band size : 286 kDa
    • All lanes : Anti-LRRK2 antibody [UDD3 30(12)] (ab133518) at 1/1000 dilution (Purified)

      Lane 1 : NIH/3T3 cell lysate
      Lane 2 : RAW264.7 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 286 kDa
      Observed band size : 286 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Anti-LRRK2 antibody [UDD3 30(12)] (ab133518) at 1/10000 dilution (Purified) + LRRK2 over-expressed 293 cell lysate at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 286 kDa
      Observed band size : 286 kDa

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    References

    This product has been referenced in:
    • Bliederhaeuser C  et al. LRRK2 contributes to monocyte dysregulation in Parkinson's disease. Acta Neuropathol Commun 4:123 (2016). Human . Read more (PubMed: 27884177) »
    • Dorval V  et al. Gene and MicroRNA transcriptome analysis of Parkinson's related LRRK2 mouse models. PLoS One 9:e85510 (2014). IP ; Mouse . Read more (PubMed: 24427314) »

    See all 6 Publications for this product

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