Overview

  • Product name
  • Description
    Rabbit polyclonal to Parkin
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Monkey
  • Immunogen

    Synthetic peptide:

    RILGEEQYTRYQQYGAEEC

    conjugated to KLH, corresponding to amino acids 304-322 of Mouse Parkin.

  • Positive control
    • WB: HEK293, SH-SY5Y and COS7 cell lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab15954 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-Fr Use at an assay dependent concentration. PubMed: 18330925
ICC/IF 1/1000. Fixation in 4% paraformaldehyde in PBS for 20 minutes at room temperature is recommended, followed by permeabilization of fixed cells with 0.1% Triton X-100 in PBS for 10 minutes at room temperature.
WB 1/1000 - 1/2000. Detects a band of approximately 52 kDa (predicted molecular weight: 51.6 kDa).
IP 1/200.

Target

  • Function
    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.
  • Tissue specificity
    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).
  • Pathway
    Protein modification; protein ubiquitination.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the RBR family. Parkin subfamily.
    Contains 1 IBR-type zinc finger.
    Contains 2 RING-type zinc fingers.
    Contains 1 ubiquitin-like domain.
  • Domain
    The ubiquitin-like domain binds the PSMD4 subunit of 26S proteasomes.
  • Post-translational
    modifications
    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.
  • Cellular localization
    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
  • Database links
  • Alternative names
    • 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

Anti-Parkin antibody images

  • Glial cell from cortical neuronal cultures co-stained with anti-parkin (red), anti-tubulin (green) and TO-PRO-3 (blue, for DNA).
  • All lanes : Anti-Parkin antibody (ab15954) at 1 µg/ml

    Lane 1 : Brain (Rat) Tissue Lysate (ab7942)
    Lane 2 : Skeletal Muscle (Rat) Tissue Lysate - normal tissue (ab29376)
    Lane 3 : Heart (Rat) Tissue Lysate (ab7940)
    Lane 4 : Human brain tissue lysate - total protein (ab29466)
    Lane 5 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 6 : Human heart tissue lysate - total protein (ab29431)
    Lane 7 : Brain (Mouse) Tissue Lysate (ab27253)
    Lane 8 : Mouse skeletal muscle tissue lysate - total protein (ab29711)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Predicted band size : 51.6 kDa
  • All lanes : Anti-Parkin antibody (ab15954) at 1/2000 dilution

    Lane 1 : Mouse hepatocytes whole cell lysate
    Lane 2 : Mouse liver whole tissue lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 51.6 kDa
    Observed band size : 52 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-Parkin antibody (ab15954)

This product has been referenced in:
  • Gao H  et al. Nur77 exacerbates PC12 cellular injury in vitro by aggravating mitochondrial impairment and endoplasmic reticulum stress. Sci Rep 6:34403 (2016). ICC/IF ; Rat . Read more (PubMed: 27679973) »
  • Im E  et al. Covalent ISG15 conjugation positively regulates the ubiquitin E3 ligase activity of parkin. Open Biol 6:N/A (2016). WB . Read more (PubMed: 27534820) »

See all 29 Publications for this product

Product Wall

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293T)
Permeabilization
Yes - 0.1% Triton X-100
Specification
HEK293T
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
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Submitted Jul 14 2017

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Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Aug 08 2016

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Application
Western blot
Sample
Mouse Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
Cardiomyocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Aug 05 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Zebrafish Tissue lysate - other (Zebrafish Brain Lysate)
Gel Running Conditions
Reduced Denaturing (Ran a 12% Precise Protein gel (12 well) (Thermo Scientific Precast Gel)
Loading amount
50 µg
Specification
Zebrafish Brain Lysate
Blocking step
Licor Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C
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Submitted Nov 06 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (Hela, MCF7, SKOV3 and CP20 cells)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
Hela, MCF7, SKOV3 and CP20 cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 4°C
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Submitted May 25 2015

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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Submitted Mar 18 2015

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Mouse Cell lysate - whole cell (embryonic fibro blast)
Specification
embryonic fibro blast
Treatment
sirna for 48h
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: rt°C
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Submitted Mar 18 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Dec 05 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Dec 05 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (hepatocytes, liver tissue)
Specification
hepatocytes, liver tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Oct 06 2014

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