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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-LRRK2 抗体 [MJFF5 (68-7)] (ab181386)

Reviews (1) Submit a question References (4)
Western blot - Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)
  • Western blot - Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)
  • Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [MJFF5 (68-7)] to LRRK2
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-LRRK2 antibody [MJFF5 (68-7)]
    LRRK2 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [MJFF5 (68-7)] to LRRK2

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WBmore details

  • 種交差性

    交差種: Mouse, Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • Human brain cortex and mouse brain cortex lysates.

  • 特記事項

    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 Epitomics, Inc., 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.

    This antibody clone (c68-7) has been re-introduced to the Epitomics catalog as characterization efforts have indicated that it performs very well at immunoprecipitating LRRK2 protein from cell lines and tissue lysate. This may facilitate investigators’ ability to more broadly examine alterations in LRRK2 protein levels and activity to interrogate changes in the biological samples.

    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.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

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

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

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

  • バッファー

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituent: 99% PBS
  • Concentration information loading...

  • 精製度

    Protein A purified

  • ポリ/モノ

    モノクローナル

  • クローン名

    MJFF5 (68-7)

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab181386の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 286 kDa.

We recommend customers use 3% milk as the blocking agent for ab181386.
特記事項
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 286 kDa.

We recommend customers use 3% milk as the blocking agent for ab181386.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    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.

  • 関連疾患

    Parkinson disease 8

  • 配列類似性

    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.

  • ドメイン

    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.

  • 翻訳後修飾

    Autophosphorylated.

  • 細胞内局在

    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

  • 参照データベース

    • 別名

      • 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

    画像

    • Western blot - Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)
      Western blot - Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: LRRK2 knockout HAP1 cell lysate (20 µg) 
      Lane 3: MEF cell lysate (20 µg)
      Lane 4: LRRK2 knockout MEF cell lysate (20 µg) 
      Lane 5: A549 cell lysate (20ug)
      Lane 6: LRRK2 knockout A549 cell lysate (20 µg) 

      Lanes 1 - 6: Merged signal (red and green). Green - ab181386 observed at 238 kDa. Red - loading control, ab18058, observed at 124 kDa.

      ab181386 was shown to specifically react with wildtype HAP1, MEF and A549 cell lysates with additional cross-reactive bands. No bands were observed when LRRK2 knockout samples were used. Wild-type and LRRK2 knockout samples were subjected to SDS-PAGE. Membranes were blocked in 3% milk for 1 hour at room temperature. ab181386 and ab130007 (loading control to Vinculin) were diluted at 2.5 ug/mL and 1/10000 respectively and incubated overnight at 4°C. 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 a generous gift from Professor Dario Alessi, MRC Protein Phosphorylation and Ubiquitination Unit (University of Dundee).

    • Western blot - Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)
      Western blot - Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)Data provided by Dr. Andrew West (University of Alabama at Birmingham)
      All lanes : Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386) at 1 µg/ml

      Lanes 1-2 : Human brain cortex lysate
      Lane 3 : mouse brain cortex lysate
      Lane 4 : LRRK2-knock-out mouse brain cortex lysate

      Developed using the ECL technique.

      Predicted band size: 286 kDa

    • Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)
      Anti-LRRK2 antibody [MJFF5 (68-7)] (ab181386)

    参考文献 (4)

    ab181386 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab181386 は 4 報の論文で使用されています。

    • Snead DM  et al. Structural basis for Parkinson's disease-linked LRRK2's binding to microtubules. Nat Struct Mol Biol 29:1196-1207 (2022). PubMed: 36510024
    • Bliederhaeuser C  et al. LRRK2 contributes to monocyte dysregulation in Parkinson's disease. Acta Neuropathol Commun 4:123 (2016). Human . PubMed: 27884177
    • Choi I  et al. LRRK2 G2019S mutation attenuates microglial motility by inhibiting focal adhesion kinase. Nat Commun 6:8255 (2015). IP ; Mouse . PubMed: 26365310
    • 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, Human . PubMed: 23560750

    レビューと Q&A

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    Flow Cytometry abreview for Anti-LRRK2 antibody [MJFF5 (68-7)]

     Excellent
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (Peripheral Blood (Monocytes))
    Permeabilization
    Yes - ‘Two-step protocol for intracellular (cytoplasmic) proteins’ from eBioscience
    Gating Strategy
    CD16+/- and CD14++/dim monocytes
    Specification
    Peripheral Blood (Monocytes)
    Preparation
    Cell harvesting/tissue preparation method: Peripheral venous blood was collected with a Monovette™ blood drawing system (Sarstedt) and processed within 3 hours post collection. Li-Heparin was used as anticoagulant. Red-blood cells (RBC) were lysed, two times washed in PBS + 10 % FCS, and the remaining leukocytes were blocked with Fc-block (1:20, BioLegend 20 min). Cell surface staining was performed with anti-human antibodies against CD14 and CD16. For intracellular LRRK2 staining the ‘Two-step protocol for intracellular (cytoplasmic) proteins’ from eBioscience was followed.
    Sample buffer: PBS + 10%FCS, Permeabilisation Buffer (eBioscience)
    Fixation
    IC Fixaction Buffer (eBioscience)
    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.

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    投稿 Dec 22 2016

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