重组Anti-PINK1抗体[EPR20730] (ab216144)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20730] to PINK1
- Suitable for: WB, ICC/IF, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-PINK1抗体[EPR20730]
参阅全部 PINK1 一抗 -
描述
兔单克隆抗体[EPR20730] to PINK1 -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IPmore details -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa cells (+/- treatment with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate; human PINK1 recombinant protein (aa156-507) . ICC/IF: HeLa cells treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours. IP: HeLA cells (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate.
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常规说明
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR20730 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab216144于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (3) |
1/1000. Predicted molecular weight: 63 kDa.
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ICC/IF | (1) |
1/500.
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IP |
1/30.
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说明 |
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WB
1/1000. Predicted molecular weight: 63 kDa. |
ICC/IF
1/500. |
IP
1/30. |
靶标
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功能
Protects against mitochondrial dysfunction during cellular stress, potentially by phosphorylating mitochondrial proteins. Involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). It is necessary for PARK2 recruitement to dysfunctional mitochondria to initiate their degradation. -
组织特异性
Highly expressed in heart, skeletal muscle and testis, and at lower levels in brain, placenta, liver, kidney, pancreas, prostate, ovary and small intestine. Present in the embryonic testis from an early stage of development. -
疾病相关
Defects in PINK1 are the cause of Parkinson disease type 6 (PARK6) [MIM:605909]. A neurodegenerative disorder characterized by parkinsonian signs such as rigidity, resting tremor and bradykinesia. A subset of patients manifest additional symptoms including hyperreflexia, autonomic instability, dementia and psychiatric disturbances. Symptoms show diurnal fluctuation and can improve after sleep. -
序列相似性
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.
Contains 1 protein kinase domain. -
翻译后修饰
Autophosphorylated. -
细胞定位
Mitochondrion outer membrane. Cytoplasm > cytosol. - Information by UniProt
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数据库链接
- Entrez Gene: 65018 Human
- NCBI: 14165272 Human
- Omim: 608309 Human
- SwissProt: Q9BXM7 Human
- Unigene: 389171 Human
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别名
- BRPK antibody
- FLJ27236 antibody
- mitochondrial antibody
see all
图片
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All lanes : Anti-PINK1 antibody [EPR20730] (ab216144) at 1/200 dilution
Lane 1 : Wild-type HEK-293 Vehicle Control CCCP, ab141229 (0 µM, 24h) cell lysate
Lane 2 : Wild-type HEK-293 Treated CCCP, ab141229 (10 µM, 24 h) cell lysate
Lane 3 : PINK1 knockout HEK-293 Vehicle Control CCCP, ab141229 (0 µM, 24 h) cell lysate
Lane 4 : PINK1 knockout HEK-293 Treated CCCP, ab141229 (10 µM, 24 h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 63 kDaAnti-PINK1 antibody [EPR20730] (ab216144) staining at 1/200 dilution shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. ab216144 was shown to bind specifically to PINK1. A band was observed at 60 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in PINK1 knockout cell line ab266393 (knockout cell lysate ab257030). Membranes were blocked in 5 % milk in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T and incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Anti-PINK1 antibody [EPR20730] (ab216144) at 1/1000 dilution + Human PINK1 recombinant protein (aa156-507), 10 ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG H&L (HRP))
Developed using the ECL technique.
Predicted band size: 63 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondDilution/blocking buffer: 5% NFDM/TBST
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Immunofluorescent analysis of 4 % paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma)(+/- treatment with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) cells labeling PINK1 with ab216144 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells treated with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
The negative controls are as follows:
-ve control: PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
All lanes : Anti-PINK1 antibody [EPR20730] (ab216144) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa cells (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 63 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking and dilution buffer: 5% NFDM/TBST
PINK1 can be induced by CCCP treatment (PMID: 24184327).
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PINK1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP. ab141229) for 24 hours) whole cell lysate with ab216144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216144 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.
Lane 1: HeLa (CCCP-treated, ab141229) lysate 10 μg (Input).
Lane 2: ab216144 IP in HeLa (CCCP-treated, ab141229) lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216144 in HeLa (CCCP-treated, ab141229) whole cell lysate.Blocking and dilution buffer: 5% NFDM/TBST.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (14)
ab216144 被引用在 14 文献中.
- Wang L et al. Puerarin inhibits FUNDC1-mediated mitochondrial autophagy and CSE-induced apoptosis of human bronchial epithelial cells by activating the PI3K/AKT/mTOR signaling pathway. Aging (Albany NY) 14:1253-1264 (2022). PubMed: 35134750
- Yu L et al. HIF-1α Alleviates High-Glucose-Induced Renal Tubular Cell Injury by Promoting Parkin/PINK1-Mediated Mitophagy. Front Med (Lausanne) 8:803874 (2021). PubMed: 35186974
- Ma KY et al. Parkinson's disease-associated VPS35 mutant reduces mitochondrial membrane potential and impairs PINK1/Parkin-mediated mitophagy. Transl Neurodegener 10:19 (2021). PubMed: 34127073
- Wang N et al. The Combination of β-Asarone and Icariin Inhibits Amyloid-β and Reverses Cognitive Deficits by Promoting Mitophagy in Models of Alzheimer's Disease. Oxid Med Cell Longev 2021:7158444 (2021). PubMed: 34887998
- Ganapathy S et al. Transport and Toxicity of Methylmercury-Cysteine in Cultured BeWo Cells. Int J Mol Sci 23:N/A (2021). PubMed: 35008820