製品の概要

  • 製品名
    Anti-Parkin antibody [PRK8]
    Parkin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [PRK8] to Parkin
  • 特異性
    Ab77924 has not been batch tested in IF and we can not guarantee that it will work in this application. However, some customers have successfully used ab77924 in IF. Please contact Abcam Scientific Support for more information.
  • アプリケーション
    適用あり: IHC-Fr, IHC-P, Flow Cyt, WB, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Drosophila C virus
  • 免疫原

    Recombinant full length Human Parkin

  • エピトープ
    The epitope is the second ring domain (aa 399-465).
  • ポジティブ・コントロール
    • This antibody gave a positive signal in SHSY-5Y whole cell lysate and in human, mouse and rat brain tissue lysates. Flow Cytometry: SHSY-5Y cells.
  • 特記事項

    This antibody clone is manufactured by Abcam.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab77924 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-Fr Use at an assay dependent concentration. PubMed: 20689587
IHC-P Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 5 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 52 kDa).

Abcam recommends using 1-3% Milk as the blocking agent.  Higher percentage blocking solutions may not give optimal results.

IP Use at an assay dependent concentration.

ターゲット情報

  • 機能
    Functions within a multiprotein E3 ubiquitin ligase complex, catalyzing the covalent attachment of ubiquitin moieties onto substrate proteins, such as BCL2, SYT11, CCNE1, GPR37, STUB1, a 22 kDa O-linked glycosylated isoform of SNCAIP, SEPT5, ZNF746 and AIMP2. Mediates monoubiquitination as well as 'Lys-48'-linked and 'Lys-63'-linked polyubiquitination of substrates depending on the context. Participates in the removal and/or detoxification of abnormally folded or damaged protein by mediating 'Lys-63'-linked polyubiquitination of misfolded proteins such as PARK7: 'Lys-63'-linked polyubiquitinated misfolded proteins are then recognized by HDAC6, leading to their recruitment to aggresomes, followed by degradation. Mediates 'Lys-63'-linked polyubiquitination of SNCAIP, possibly playing a role in Lewy-body formation. Mediates monoubiquitination of BCL2, thereby acting as a positive regulator of autophagy. Promotes the autophagic degradation of dysfunctional depolarized mitochondria. Mediates 'Lys-48'-linked polyubiquitination of ZNF746, followed by degradation of ZNF746 by the proteasome; possibly playing a role in role in regulation of neuron death. Limits the production of reactive oxygen species (ROS). Loss of this ubiquitin ligase activity appears to be the mechanism underlying pathogenesis of PARK2. May protect neurons against alpha synuclein toxicity, proteasomal dysfunction, GPR37 accumulation, and kainate-induced excitotoxicity. May play a role in controlling neurotransmitter trafficking at the presynaptic terminal and in calcium-dependent exocytosis. Regulates cyclin-E during neuronal apoptosis. May represent a tumor suppressor gene.
  • 組織特異性
    Highly expressed in the brain including the substantia nigra. Expressed in heart, testis and skeletal muscle. Expression is down-regulated or absent in tumor biopsies, and absent in the brain of PARK2 patients. Overexpression protects dopamine neurons from kainate-mediated apoptosis. Found in serum (at protein level).
  • パスウェイ
    Protein modification; protein ubiquitination.
  • 関連疾患
    Defects in PARK2 are a cause of Parkinson disease (PARK) [MIM:168600]. A complex neurodegenerative disorder characterized by bradykinesia, resting tremor, muscular rigidity and postural instability. Additional features are characteristic postural abnormalities, dysautonomia, dystonic cramps, and dementia. The pathology of Parkinson disease involves the loss of dopaminergic neurons in the substantia nigra and the presence of Lewy bodies (intraneuronal accumulations of aggregated proteins), in surviving neurons in various areas of the brain. The disease is progressive and usually manifests after the age of 50 years, although early-onset cases (before 50 years) are known. The majority of the cases are sporadic suggesting a multifactorial etiology based on environmental and genetic factors. However, some patients present with a positive family history for the disease. Familial forms of the disease usually begin at earlier ages and are associated with atypical clinical features.
    Defects in PARK2 are the cause of Parkinson disease type 2 (PARK2) [MIM:600116]; also known as early-onset parkinsonism with diurnal fluctuation (EPDF) or autosomal recessive juvenile Parkinson disease (PDJ). A neurodegenerative disorder characterized by bradykinesia, rigidity, postural instability, tremor, and onset usually befor 40. It differs from classic Parkinson disease by early DOPA-induced dyskinesia, diurnal fluctuation of the symptoms, sleep benefit, dystonia and hyper-reflexia. Dementia is absent. Pathologically, patients show loss of dopaminergic neurons in the substantia nigra, similar to that seen in Parkinson disease; however, Lewy bodies (intraneuronal accumulations of aggregated proteins) are absent.
    Note=Defects in PARK2 may be involved in the development and/or progression of ovarian cancer.
  • 配列類似性
    Belongs to the RBR family. Parkin subfamily.
    Contains 1 IBR-type zinc finger.
    Contains 2 RING-type zinc fingers.
    Contains 1 ubiquitin-like domain.
  • ドメイン
    The ubiquitin-like domain binds the PSMD4 subunit of 26S proteasomes.
  • 翻訳後修飾
    Auto-ubiquitinates in an E2-dependent manner leading to its own degradation. Also polyubiquitinated by RNF41 for proteasomal degradation.
    S-nitrosylated. The inhibition of PARK2 ubiquitin E3 ligase activity by S-nitrosylation could contribute to the degenerative process in PD by impairing the ubiquitination of PARK2 substrates.
  • 細胞内局在
    Cytoplasm > cytosol. Nucleus. Endoplasmic reticulum. Mitochondrion. Mainly localizes in the cytosol. Co-localizes with SYT11 in neutrites. Co-localizes with SNCAIP in brainstem Lewy bodies. Relocates to dysfunctional mitochondria that have lost the mitochondial membrane potential; recruitement to mitochondria is PINK1-dependent.
  • Information by UniProt
  • 参照データベース
  • 別名
    • AR JP antibody
    • E3 ubiquitin ligase antibody
    • E3 ubiquitin protein ligase parkin antibody
    • E3 ubiquitin-protein ligase parkin antibody
    • FRA6E antibody
    • LPRS 2 antibody
    • LPRS2 antibody
    • PARK 2 antibody
    • Park2 antibody
    • Parkin 2 antibody
    • Parkinson disease (autosomal recessive juvenile) 2 antibody
    • Parkinson disease (autosomal recessive, juvenile) 2, parkin antibody
    • Parkinson disease protein 2 antibody
    • Parkinson juvenile disease protein 2 antibody
    • Parkinson protein 2 E3 ubiquitin protein ligase antibody
    • Parkinson protein 2, E3 ubiquitin protein ligase (parkin) antibody
    • PDJ antibody
    • PRKN 2 antibody
    • PRKN antibody
    • PRKN2 antibody
    • PRKN2_HUMAN antibody
    • Ubiquitin E3 ligase PRKN antibody
    see all

画像

  • All lanes : Anti-Parkin antibody [PRK8] (ab77924) at 1/2000 dilution

    Lane 1 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate
    Lane 4 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 52 kDa
    Additional bands at : 55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes

    All lanes blocked with 3% milk.

  • Overlay histogram showing SH-SY5Y cells stained with ab77924 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab77924, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunohistochemical analysis of transgenic Zebrafish expressing Human Parkin, staining Parkin with ab77924.

    Tissue was fixed with paraformaldehyde and blocked with 10% newborn calf serum with 0.1% Tween for 2 hours. Samples were incubated with primary antibody (1/200) overnight at 4°C. An AlexaFluor®488-conjugated anti-mouse IgG was used as the secondary antibody.

参考文献

This product has been referenced in:
  • Lieberknecht V  et al. Pramipexole, a Dopamine D2/D3 Receptor-Preferring Agonist, Prevents Experimental Autoimmune Encephalomyelitis Development in Mice. Mol Neurobiol 54:1033-1045 (2017). Read more (PubMed: 26801190) »
  • Wei L  et al. Involvement of PINK1/parkin-mediated mitophagy in ZnO nanoparticle-induced toxicity in BV-2 cells. Int J Nanomedicine 12:1891-1903 (2017). Read more (PubMed: 28331313) »

See all 17 Publications for this product

レビューと Q&A

Thank you for your response.
As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily off...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (liver)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Verified customer

投稿 Aug 02 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Chinese Hamster Cell (CHO cell)
Specification
CHO cell
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Paraformaldehyde
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投稿 Mar 18 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (liver, hepatocytes)
Specification
liver, hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

投稿 Jan 07 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - other (brain mitochondria)
Loading amount
40 µg
Specification
brain mitochondria
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

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Verified customer

投稿 Feb 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Drosophila C Virus Tissue lysate - whole (fly heart tube)
Loading amount
10 µg
Specification
fly heart tube
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

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Verified customer

投稿 Feb 11 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Drosophila C virus Cell (fly heart tube)
Permeabilization
Yes - 1% triton x100
Specification
fly heart tube
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Fixative
Paraformaldehyde
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Verified customer

投稿 Feb 08 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (mouse heart)
Total protein in input
200 µg
Specification
mouse heart
Immuno-precipitation step
Protein G
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Verified customer

投稿 Feb 08 2013

Vielen Dank nochmal für Ihre Anfrage.



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