Anti-Parkin antibody [PRK8] (ab77924) is a mouse monoclonal antibody detecting Parkin in Western Blot. Suitable for Human, Mouse, Rat.
- KO validated for confirmed specificity
- Over 150 publications
- Trusted since 2009
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Flow Cyt | WB | |
---|---|---|
Human | Not recommended | Tested |
Mouse | Not recommended | Tested |
Rat | Not recommended | Tested |
Drosophila melanogaster | Not recommended | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Human, Drosophila melanogaster | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/2000 | Notes Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
Species Rat | Dilution info 1/1000 - 1/2000 | Notes Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
Species Human | Dilution info 1/1000 - 1/2000 | Notes Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
Species | Dilution info | Notes |
---|---|---|
Species Drosophila melanogaster | Dilution info - | Notes - |
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Functions within a multiprotein E3 ubiquitin ligase complex, catalyzing the covalent attachment of ubiquitin moieties onto substrate proteins (PubMed:10888878, PubMed:10973942, PubMed:11431533, PubMed:12150907, PubMed:12628165, PubMed:15105460, PubMed:16135753, PubMed:21376232, PubMed:21532592, PubMed:22396657, PubMed:23620051, PubMed:23754282, PubMed:24660806, PubMed:24751536, PubMed:29311685, PubMed:32047033). Substrates include SYT11 and VDAC1 (PubMed:29311685, PubMed:32047033). Other substrates are BCL2, CCNE1, GPR37, RHOT1/MIRO1, MFN1, MFN2, STUB1, SNCAIP, SEPTIN5, TOMM20, USP30, ZNF746, MIRO1 and AIMP2 (PubMed:10888878, PubMed:10973942, PubMed:11431533, PubMed:12150907, PubMed:12628165, PubMed:15105460, PubMed:16135753, PubMed:21376232, PubMed:21532592, PubMed:22396657, PubMed:23620051, PubMed:23754282, PubMed:24660806, PubMed:24751536). Mediates monoubiquitination as well as 'Lys-6', 'Lys-11', 'Lys-48'-linked and 'Lys-63'-linked polyubiquitination of substrates depending on the context (PubMed:19229105, PubMed:20889974, PubMed:25474007, PubMed:25621951, PubMed:32047033). 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 (PubMed:17846173, PubMed:19229105). Mediates 'Lys-63'-linked polyubiquitination of a 22 kDa O-linked glycosylated isoform of SNCAIP, possibly playing a role in Lewy-body formation (PubMed:11431533, PubMed:11590439, PubMed:15105460, PubMed:15728840, PubMed:19229105). Mediates monoubiquitination of BCL2, thereby acting as a positive regulator of autophagy (PubMed:20889974). Protects against mitochondrial dysfunction during cellular stress, by acting downstream of PINK1 to coordinate mitochondrial quality control mechanisms that remove and replace dysfunctional mitochondrial components (PubMed:11439185, PubMed:18957282, PubMed:19029340, PubMed:19966284, PubMed:21376232, PubMed:22082830, PubMed:22396657, PubMed:23620051, PubMed:23933751, PubMed:24660806, PubMed:24784582, PubMed:24896179, PubMed:25474007, PubMed:25527291, PubMed:32047033). Depending on the severity of mitochondrial damage and/or dysfunction, activity ranges from preventing apoptosis and stimulating mitochondrial biogenesis to regulating mitochondrial dynamics and eliminating severely damaged mitochondria via mitophagy (PubMed:11439185, PubMed:19029340, PubMed:19801972, PubMed:19966284, PubMed:21376232, PubMed:22082830, PubMed:22396657, PubMed:23620051, PubMed:23685073, PubMed:23933751, PubMed:24896179, PubMed:25527291, PubMed:32047033, PubMed:33499712). Activation and recruitment onto the outer membrane of damaged/dysfunctional mitochondria (OMM) requires PINK1-mediated phosphorylation of both PRKN and ubiquitin (PubMed:24660806, PubMed:24784582, PubMed:25474007, PubMed:25527291). After mitochondrial damage, functions with PINK1 to mediate the decision between mitophagy or preventing apoptosis by inducing either the poly- or monoubiquitination of VDAC1, respectively; polyubiquitination of VDAC1 promotes mitophagy, while monoubiquitination of VDAC1 decreases mitochondrial calcium influx which ultimately inhibits apoptosis (PubMed:27534820, PubMed:32047033). When cellular stress results in irreversible mitochondrial damage, promotes the autophagic degradation of dysfunctional depolarized mitochondria (mitophagy) by promoting the ubiquitination of mitochondrial proteins such as TOMM20, RHOT1/MIRO1, MFN1 and USP30 (PubMed:19029340, PubMed:19966284, PubMed:21753002, PubMed:22396657, PubMed:23620051, PubMed:23685073, PubMed:23933751, PubMed:24896179, PubMed:25527291). Preferentially assembles 'Lys-6'-, 'Lys-11'- and 'Lys-63'-linked polyubiquitin chains, leading to mitophagy (PubMed:25621951, PubMed:32047033). The PINK1-PRKN pathway also promotes fission of damaged mitochondria by PINK1-mediated phosphorylation which promotes the PRKN-dependent degradation of mitochondrial proteins involved in fission such as MFN2 (PubMed:23620051). This prevents the refusion of unhealthy mitochondria with the mitochondrial network or initiates mitochondrial fragmentation facilitating their later engulfment by autophagosomes (PubMed:23620051). Regulates motility of damaged mitochondria via the ubiquitination and subsequent degradation of MIRO1 and MIRO2; in motor neurons, this likely inhibits mitochondrial intracellular anterograde transport along the axons which probably increases the chance of the mitochondria undergoing mitophagy in the soma (PubMed:22396657). Involved in mitochondrial biogenesis via the 'Lys-48'-linked polyubiquitination of transcriptional repressor ZNF746/PARIS which leads to its subsequent proteasomal degradation and allows activation of the transcription factor PPARGC1A (PubMed:21376232). Limits the production of reactive oxygen species (ROS) (PubMed:18541373). Regulates cyclin-E during neuronal apoptosis (PubMed:12628165). In collaboration with CHPF isoform 2, may enhance cell viability and protect cells from oxidative stress (PubMed:22082830). Independently of its ubiquitin ligase activity, protects from apoptosis by the transcriptional repression of p53/TP53 (PubMed:19801972). May protect neurons against alpha synuclein toxicity, proteasomal dysfunction, GPR37 accumulation, and kainate-induced excitotoxicity (PubMed:11439185). May play a role in controlling neurotransmitter trafficking at the presynaptic terminal and in calcium-dependent exocytosis. May represent a tumor suppressor gene (PubMed:12719539).
PARK2, PRKN, E3 ubiquitin-protein ligase parkin, Parkin, Parkin RBR E3 ubiquitin-protein ligase, Parkinson juvenile disease protein 2, Parkinson disease protein 2
Anti-Parkin antibody [PRK8] (ab77924) is a mouse monoclonal antibody detecting Parkin in Western Blot. Suitable for Human, Mouse, Rat.
- KO validated for confirmed specificity
- Over 150 publications
- Trusted since 2009
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
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This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Terms & Conditions.
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 Human PARK2 (Parkin) knockout SH-SY5Y cell line ab280042 (PRKN knockout cell lysate Human PARK2 (Parkin) knockout SH-SY5Y 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 Anti-GAPDH antibody [EPR16891] - Loading Control 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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-Parkin antibody [PRK8] (ab77924) at 5 µg/mL
Lane 1: Wild-type SH-SY5Y cell lysate at 20 µg
Lane 2: PRKN knockout SH-SY5Y cell lysate at 20 µg
Lane 2: Western blot - Human PARK2 (Parkin) knockout SH-SY5Y cell line (Human PARK2 (Parkin) knockout SH-SY5Y cell line ab280042)
Lane 3: Human Brain tissue lysate at 20 µg
Lane 4: HUVEC cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 49 kDa
All lanes: Western blot - Anti-Parkin antibody [PRK8] (ab77924) at 5 µg/mL
Lane 1: Human brain tissue lysate at 20 µg
Lane 2: Mouse brain tissue lysate at 20 µg
Lane 3: Rat brain tissue lysate at 20 µg
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: 8min
All lanes blocked with 3% milk.
All lanes: Western blot - Anti-Parkin antibody [PRK8] (ab77924) at 1/2000 dilution
Lane 1: SH-SY5Y (Human neuroblastoma cell line) Whole Cell Lysate at 20 µg
Lane 2: Brain (Rat) Tissue Lysate at 20 µg
Lane 3: Brain (Mouse) Tissue Lysate at 20 µg
Lane 4: Brain (Human) Tissue Lysate at 20 µg
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
Exposure time: 20min
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