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  1. Link

    products/primary-antibodies/parkin-antibody-prk8-ab77924.pdf

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Neuroscience Neurology process Neurodegenerative disease Parkinson's disease Parkin / PARK
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Validated using a knockout cell line

Anti-Parkin antibody [PRK8] (ab77924)

  • Datasheet
  • SDS
Reviews (12) Submit a question References (101)

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

Key features and details

  • Mouse monoclonal [PRK8] to Parkin
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG2b

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Knockout
Product image
Human PARK2 (Parkin) knockout SH-SY5Y cell line (ab280042)

View more associated products

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: WBmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: 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 and HUVEC whole cell lysate. Human, mouse and rat brain tissue lysates.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, 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
    • Neuroscience
    • Diseases
    • Neuroscience
    • Processes

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 (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

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

    • Recombinant Human Parkin protein (ab140806)

Applications

The Abpromise guarantee

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
WB (7)
1/2000. 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.

Notes
WB
1/2000. 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.

Application notes
Is unsuitable for Flow Cyt.

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 5 µg/ml

    Lane 1 : Wild-type SH-SY5Y cell lysate
    Lane 2 : PRKN knockout SH-SY5Y cell lysate
    Lane 3 : Human Brain tissue lysate
    Lane 4 : HUVEC cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 49 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab77924 observed at 49 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

    ab77924 was shown to react with Parkin in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PRKN knockout cell line ab280042 (PRKN knockout cell lysate ab280101). Wild-type SH-SY5Y and PRKN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab77924 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-Parkin antibody [PRK8] (ab77924)
    Western blot - Anti-Parkin antibody [PRK8] (ab77924)
    All lanes : Anti-Parkin antibody [PRK8] (ab77924) at 5 µg/ml

    Lane 1 : Human brain tissue lysate
    Lane 2 : Mouse brain tissue lysate
    Lane 3 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Predicted band size: 52 kDa
    Observed band size: 52 kDa


    Exposure time: 8 minutes


    Blocking buffer: 3% milk.

  • 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 : Brain (Human) Tissue Lysate

    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
    Observed band size: 55 kDa why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    All lanes blocked with 3% milk.

Protocols

  • Mouse on Mouse staining protocol

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (101)

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

ab77924 has been referenced in 101 publications.

  • Choi GE  et al. BNIP3L/NIX-mediated mitophagy protects against glucocorticoid-induced synapse defects. Nat Commun 12:487 (2021). PubMed: 33473105
  • Zhou B  et al. Mitochondrial quality control protects photoreceptors against oxidative stress in the H2O2-induced models of retinal degeneration diseases. Cell Death Dis 12:413 (2021). PubMed: 33879768
  • Liu D  et al. c-MYC-induced long noncoding RNA MEG3 aggravates kidney ischemia-reperfusion injury through activating mitophagy by upregulation of RTKN to trigger the Wnt/ß-catenin pathway. Cell Death Dis 12:191 (2021). PubMed: 33602903
  • Roca-Agujetas V  et al. Cholesterol alters mitophagy by impairing optineurin recruitment and lysosomal clearance in Alzheimer's disease. Mol Neurodegener 16:15 (2021). PubMed: 33685483
  • Castro-Gonzalez S  et al. HIV-1 Nef counteracts autophagy restriction by enhancing the association between BECN1 and its inhibitor BCL2 in a PRKN-dependent manner. Autophagy 17:553-577 (2021). PubMed: 32097085
View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

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
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. Tom Ryan

Verified customer

Submitted Sep 29 2020

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
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 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
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 Jul 24 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
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. 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
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 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
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 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
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 Feb 11 2013

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