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  1. Link

    lrrk2-antibody-mjff2-c41-2-ab133474.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 [MJFF2 (c41-2)] (ab133474)

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  • SDS
Reviews (2)Q&A (1)References (86)

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Immunocytochemistry/ Immunofluorescence - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
  • Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
  • Immunocytochemistry/ Immunofluorescence - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
  • Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
  • Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
  • Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
  • Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [MJFF2 (c41-2)] to LRRK2
  • Suitable for: ICC/IF, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free HRP

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Overview

  • Product name

    Anti-LRRK2 antibody [MJFF2 (c41-2)]
    See all LRRK2 primary antibodies
  • Description

    Rabbit monoclonal [MJFF2 (c41-2)] to LRRK2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IPmore 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.
    Database link: Q5S007

  • Positive control

    • WB: A549 and MEF cell lysates; HEK293 cells transfected with LRRK2 cell lysate. ICC/IF: Neuro-2a cells. IHC-P: SNc of 1 y-old Rgs6-/- mice; Mouse brain tissue IP: Mouse cerebral cortex, A549 .
  • 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]

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    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.

    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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    MJFF2 (c41-2)
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-LRRK2 antibody [MJFF2 (c41-2)] - BSA and Azide free (ab172378)
    • HRP Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab195024)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Anti-IKB alpha antibody [EP697] - BSA and Azide free (ab183134)
  • Isotype control

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

    • MLi-2, LRRK2 inhibitor (ab254528)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133474 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
1/50.
WB (2)
1/10000 - 1/50000. Predicted molecular weight: 286 kDa.
IP
Use at an assay dependent concentration. (2-5 µg)
Notes
ICC/IF
1/50.
WB
1/10000 - 1/50000. Predicted molecular weight: 286 kDa.
IP
Use at an assay dependent concentration. (2-5 µg)

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

    • Immunocytochemistry/ Immunofluorescence - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
      Immunocytochemistry/ Immunofluorescence - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)Rassu, M. et al PLoS One. 2017 Jun 5;12(6):e0179082. doi: 10.1371/journal.pone.0179082. eCollection 2017. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Evaluation of DRD1 intracellular and extracellular level by BPA upon dopamine treatment in SH-SY5Y-DRD1 cells untransduced or transduced by WT or G2019S LRRK2

      (C and D) DRD1 localization at basal conditions (C) and upon 15 minutes (D) of dopamine treatment of SH-SY5Y-DRD1 cells transduced or not by alpha-synuclein WT or A53T. After agonist treatment the cells were fixed and incubated with the different primary antibodies (anti-FLAG for DRD1 and ab133474) and with Alexa647-conjugated secondary antibody (red) or Alexa488-conjugated secondary antibody (green) for DRD1 or LRRK2 respectively. Scale bars = 10μm.

       

    • Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
      Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)

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

      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 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.

      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).

    • Immunocytochemistry/ Immunofluorescence - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
      Immunocytochemistry/ Immunofluorescence - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)

      ab133474 staining LRRK2 in Neuro-2a (mouse neuroblastoma cell line) cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/500). ab150077 was used as the secondary antibody (1/1000). Tubulin stained using ab7291 (1/1000) and ab150120 (1/1000) as the secondary. Nuclear counter stained with DAPI.

    • Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
      Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)

      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)(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 (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.

    • Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
      Immunoprecipitation - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)Stafa K. et al., PLoS Genet. 2012;8(2):e1002526. Fig 1 doi: 10.1371/journal.pgen.1002526. Reproduced under the Creative Commons license

      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.

    • Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
      Western blot - Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)This image is courtesy of Zhuohua Zhang Lab (Sanford-Burnham Medical Research Institute)
      All lanes : Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474) at 1/10000 dilution

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

    • Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)
      Anti-LRRK2 antibody [MJFF2 (c41-2)] (ab133474)

    Protocols

    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (86)

    Publishing research using ab133474? Please let us know so that we can cite the reference in this datasheet.

    ab133474 has been referenced in 86 publications.

    • De Miranda BR  et al. The industrial solvent trichloroethylene induces LRRK2 kinase activity and dopaminergic neurodegeneration in a rat model of Parkinson's disease. Neurobiol Dis 153:105312 (2021). PubMed: 33636387
    • Chen X  et al. Parkinson's disease-related Leucine-rich repeat kinase 2 modulates nuclear morphology and genomic stability in striatal projection neurons during aging. Mol Neurodegener 15:12 (2020). PubMed: 32075681
    • Herbst S  et al. LRRK2 activation controls the repair of damaged endomembranes in macrophages. EMBO J 39:e104494 (2020). PubMed: 32643832
    • Bonet-Ponce L  et al. LRRK2 mediates tubulation and vesicle sorting from lysosomes. Sci Adv 6:N/A (2020). PubMed: 33177079
    • Baptista MAS  et al. LRRK2 inhibitors induce reversible changes in nonhuman primate lungs without measurable pulmonary deficits. Sci Transl Med 12:N/A (2020). PubMed: 32321864
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-LRRK2 antibody [MJFF2 (c41-2)]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (Fetal Enteric nervous system)
    Gel Running Conditions
    Reduced Denaturing (Tris Acétate 3-8%)
    Loading amount
    30 µg
    Specification
    Fetal Enteric nervous system
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Tony Durand

    Verified customer

    Submitted Feb 23 2022

    Western blot abreview for Anti-LRRK2 antibody [MJFF2 (c41-2)]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293FT cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Treatment
    NTC and LRRK2 siRNA
    Specification
    HEK293FT cells
    Blocking step
    Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 18 2020

    Question

    ab133450, ab133474, ab133475, ab133476:

    confirmer:
    présence ou pas de BSA dans tampon de stockage
    méthode purification
    concentration
    stockage à +4ºC.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 07 2012

    Answer

    Merci de votre patience.


    Après vérification, mes collègues m'ont confirmé que les fiches techniques Epitomics sont exactes:

    ab133474, ab133475 et ab133476 sont des anticorps non purifiés (supernageants de culture). Les 3 sont sans BSA. Les concentrations de ces 3 références sont connues mais dépendent du lot. Indiquez-moi les numéros des lots que vous avez reçus et je pourrai vous communiquer la concentration de chaque produit.
    Les erreurs sur les fiches techniques Abcam de ces 3 produits vont être corrigées dans les plus brefs délais.

    Le ab133450 contient bien de la BSA. Pas d'erreur sur les fiches techniques Epitomics ou Abcam pour ce produit. Le point d'interrogation après le 4ºC provient d'un problème informatique lors de la sauvegarde de la fiche technique en pdf.




    J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Read More

    Abcam Scientific Support

    Answered on Dec 07 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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