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    parkin-antibody-prk8-ab77924.pdf

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Neuroscience Neurology process Neurodegenerative disease Parkinson's disease Parkin / PARK
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Anti-Parkin antibody [PRK8] (ab77924)

  • Datasheet
  • SDS
Reviews (12) Submit a question References (64)

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Western blot - Anti-Parkin antibody [PRK8] (ab77924)
  • Flow Cytometry - Anti-Parkin antibody [PRK8] (ab77924)

Key features and details

  • Mouse monoclonal [PRK8] to Parkin
  • Suitable for: Flow Cyt, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG2b

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Overview

  • Product name

    Anti-Parkin antibody [PRK8]
    See all Parkin primary antibodies
  • Description

    Mouse monoclonal [PRK8] to Parkin
  • Host species

    Mouse
  • Tested applications

    Suitable for: Flow Cyt, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Drosophila melanogaster
  • Immunogen

    Recombinant full length protein corresponding to Human Parkin.

  • Epitope

    The epitope is the second ring domain (aa 399-465).
  • Positive control

    • WB: SH-SY5Y whole cell lysate. Human, mouse and rat brain tissue lysates. Wild-type HAP1 whole cell lysate. Flow Cyt: SH-SY5Y cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    PRK8
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Parkinson's disease
    • Parkin / PARK
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E3 Enzymes
    • RING Finger E3 Ligase
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitophagy fission and fusion

Associated products

  • Alternative Versions

    • Anti-Parkin antibody [PRK8] - BSA and Azide free (ab264105)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • Recombinant Protein

    • Recombinant Human Parkin protein (ab140806)

Applications

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.

Application Abreviews Notes
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.

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.
  • Target information above from: UniProt accession O60260 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5071 Human
    • Entrez Gene: 50873 Mouse
    • Entrez Gene: 56816 Rat
    • Omim: 602544 Human
    • SwissProt: O60260 Human
    • SwissProt: Q9WVS6 Mouse
    • SwissProt: Q9JK66 Rat
    • Unigene: 132954 Human
    • Unigene: 311110 Mouse
    • Unigene: 207194 Rat
    see all
  • 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

Images

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

    Lane 1 : SH-SY5Y (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
    All lanes : 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.

  • Flow Cytometry - Anti-Parkin antibody [PRK8] (ab77924)
    Flow Cytometry - Anti-Parkin antibody [PRK8] (ab77924)

    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.

Protocols

  • Mouse on Mouse staining protocol

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (64)

    Publishing research using ab77924? Please let us know so that we can cite the reference in this datasheet.

    ab77924 has been referenced in 64 publications.

    • Zheng J  et al. MSCs ameliorate hepatocellular apoptosis mediated by PINK1-dependent mitophagy in liver ischemia/reperfusion injury through AMPKa activation. Cell Death Dis 11:256 (2020). PubMed: 32312955
    • Duan C  et al. Drp1 regulates mitochondrial dysfunction and dysregulated metabolism in ischemic injury via Clec16a-, BAX-, and GSH- pathways. Cell Death Dis 11:251 (2020). PubMed: 32312970
    • Xia ML  et al. Astragaloside IV inhibits astrocyte senescence: implication in Parkinson's disease. J Neuroinflammation 17:105 (2020). PubMed: 32252767
    • Wu H  et al. TIM-4 interference in Kupffer cells against CCL4-induced liver fibrosis by mediating Akt1/Mitophagy signalling pathway. Cell Prolif 53:e12731 (2020). PubMed: 31755616
    • Zhang S  et al. HMGB1/RAGE axis mediates stress-induced RVLM neuroinflammation in mice via impairing mitophagy flux in microglia. J Neuroinflammation 17:15 (2020). PubMed: 31924219
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 12 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Parkin antibody [PRK8]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa cell transfected with HA-Parkin)
    Permeabilization
    Yes - 0.1% Triton in PBS (5 mins)
    Specification
    HeLa cell transfected with HA-Parkin
    Blocking step
    BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 06 2021

    Western blot abreview for Anti-Parkin antibody [PRK8]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    10 µg
    Specification
    HeLa
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Tom Ryan

    Verified customer

    Submitted Sep 29 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Parkin antibody [PRK8]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HTC116 human colorectal carcinoma)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    HTC116 human colorectal carcinoma
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Dimitra Kalamida

    Verified customer

    Submitted Dec 13 2019

    Western blot abreview for Anti-Parkin antibody [PRK8]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T, ARPE-19)
    Gel Running Conditions
    Reduced Denaturing (4-12% BisTris Gel)
    Loading amount
    10 µg
    Specification
    293T, ARPE-19
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 24 2019

    Western blot abreview for Anti-Parkin antibody [PRK8]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (retina, RPE)
    Gel Running Conditions
    Reduced Denaturing (4-12% BisTris Gel)
    Loading amount
    30 µg
    Specification
    retina, RPE
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 24 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Parkin antibody [PRK8]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Neural progenitor cell)
    Specification
    Neural progenitor cell
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 08 2019

    Western blot abreview for Anti-Parkin antibody [PRK8]

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - other (head and neck cancer)
    Gel Running Conditions
    Reduced Denaturing (12% SDS page gel)
    Loading amount
    30 µg
    Treatment
    5uM DIM-C-pPhtBu for 24h
    Specification
    head and neck cancer
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Dr. Sung Un Kang

    Verified customer

    Submitted Sep 18 2018

    Western blot abreview for Anti-Parkin antibody [PRK8]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 02 2016

    Western blot abreview for Anti-Parkin antibody [PRK8]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 07 2015

    Western blot abreview for Anti-Parkin antibody [PRK8]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2013

    1-10 of 12 Abreviews or Q&A

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