Anti-PARK7/DJ1抗体[EP2816Y] (ab76241)
Key features and details
- Rabbit monoclonal [EP2816Y] to PARK7/DJ1
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Related conjugates and formulations
概述
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产品名称
Anti-PARK7/DJ1抗体[EP2816Y]
参阅全部 PARK7/DJ1 一抗 -
描述
兔单克隆抗体[EP2816Y] to PARK7/DJ1 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, Flow Cyt (Intra)more details
不适用于: IP -
种属反应性
与反应: Human
预测可用于: Mouse, Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: TF-1, Jurkat, HEK293T, HAP1 and HeLa cell lysates; Human brain nuclear extract tissue lysate; Human brain tissue lysate. Flow Cyt (intra): Jurkat cells. IHC-P: Human brain tissue.
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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.
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
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Tissue culture supernatant -
克隆
单克隆 -
克隆编号
EP2816Y -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab76241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 | (1) |
1/10000 - 1/20000. Detects a band of approximately 24 kDa (predicted molecular weight: 20 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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WB
1/10000 - 1/20000. Detects a band of approximately 24 kDa (predicted molecular weight: 20 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Protects cells against oxidative stress and cell death. Plays a role in regulating expression or stability of the mitochondrial uncoupling proteins SLC25A14 and SLC25A27 in dopaminergic neurons of the substantia nigra pars compacta and attenuates the oxidative stress induced by calcium entry into the neurons via L-type channels during pacemaking. Eliminates hydrogen peroxide and protects cells against hydrogen peroxide-induced cell death. May act as an atypical peroxiredoxin-like peroxidase that scavenges hydrogen peroxide. Following removal of a C-terminal peptide, displays protease activity and enhanced cytoprotective action against oxidative stress-induced apoptosis. Stabilizes NFE2L2 by preventing its association with KEAP1 and its subsequent ubiquitination. Binds to OTUD7B and inhibits its deubiquitinating activity. Enhances RELA nuclear translocation. Binds to a number of mRNAs containing multiple copies of GG or CC motifs and partially inhibits their translation but dissociates following oxidative stress. Required for correct mitochondrial morphology and function and for autophagy of dysfunctional mitochondria. Regulates astrocyte inflammatory responses. Acts as a positive regulator of androgen receptor-dependent transcription. Prevents aggregation of SNCA. Plays a role in fertilization. Has no proteolytic activity. Has cell-growth promoting activity and transforming activity. May function as a redox-sensitive chaperone. -
组织特异性
Highly expressed in pancreas, kidney, skeletal muscle, liver, testis and heart. Detected at slightly lower levels in placenta and brain. Detected in astrocytes, Sertoli cells, spermatogonia, spermatids and spermatozoa. -
疾病相关
Defects in PARK7 are the cause of Parkinson disease type 7 (PARK7) [MIM:606324]. A neurodegenerative disorder characterized by resting tremor, postural tremor, bradykinesia, muscular rigidity, anxiety and psychotic episodes. PARK7 has onset before 40 years, slow progression and initial good response to levodopa. Some patients may show traits reminiscent of amyotrophic lateral sclerosis-parkinsonism/dementia complex (Guam disease). -
序列相似性
Belongs to the peptidase C56 family. -
翻译后修饰
Sumoylated on Lys-130 by PIAS2 or PIAS4; which is enhanced after ultraviolet irradiation and essential for cell-growth promoting activity and transforming activity.
Cys-106 is easily oxidized to sulfinic acid.
Undergoes cleavage of a C-terminal peptide and subsequent activation of protease activity in response to oxidative stress. -
细胞定位
Cytoplasm. Nucleus. Mitochondrion. Under normal conditions, located predominantly in the cytoplasm and, to a lesser extent, in the nucleus and mitochondrion. Translocates to the mitochondrion and subsequently to the nucleus in response to oxidative stress and exerts an increased cytoprotective effect against oxidative damage. Detected in tau inclusions in brains from neurodegenerative disease patients. - Information by UniProt
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数据库链接
- Entrez Gene: 11315 Human
- Entrez Gene: 57320 Mouse
- Entrez Gene: 117287 Rat
- Omim: 602533 Human
- SwissProt: Q99497 Human
- SwissProt: Q99LX0 Mouse
- SwissProt: O88767 Rat
- Unigene: 419640 Human
see all -
别名
- CAP1 antibody
- DJ-1 antibody
- DJ1 antibody
see all
图片
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All lanes : Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : PARK7 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Human brain nuclear fraction tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 20 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab76241 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
ab76241 Anti-PARK7/DJ1 antibody [EP2816Y] was shown to specifically react with PARK7/DJ1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266338 (knockout cell lysate ab257016) was used. Wild-type and PARK7/DJ1 knockout samples were subjected to SDS-PAGE. ab76241 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/10000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : PARK/DJ1 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Human brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 20 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PARK/DJ1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human brain tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab76241 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab76241 was shown to specifically react with PARK7/DJ1 in wild-type HAP1 cells. No band was observed when PARK7/DJ1 knockout samples were used. Wild-type and PARK7/DJ1 knockout samples were subjected to SDS-PAGE. ab76241 and ab8245 (loading control to PARK7/DJ1) were both diluted 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging. -
All lanes : Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/20000 dilution
Lane 1 : TF-1 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/1000 dilution
Predicted band size: 20 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted? -
Overlay histogram showing Jurkat cells stained with ab76241 (red line). The cells were fixed with 80% methanol (5 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 (ab76241, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Immunohistochemical analysis of paraffin-embedded human brain tissue using ab76241 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (8)
ab76241 被引用在 8 文献中.
- Goljanek-Whysall K et al. miR-181a regulates p62/SQSTM1, parkin, and protein DJ-1 promoting mitochondrial dynamics in skeletal muscle aging. Aging Cell 19:e13140 (2020). PubMed: 32291905
- Tsai CY et al. Redox-active DJ-1 sustains brainstem cardiovascular regulation via maintenance of mitochondrial function during mevinphos intoxication. Neurochem Int 139:104791 (2020). PubMed: 32650030
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
- Di Nottia M et al. DJ-1 modulates mitochondrial response to oxidative stress: clues from a novel diagnosis of PARK7. Clin Genet N/A:N/A (2016). PubMed: 27460976
- Prahlad J et al. Use of cysteine-reactive cross-linkers to probe conformational flexibility of human DJ-1 demonstrates that Glu18 mutations are dimers. J Neurochem 130:839-53 (2014). PubMed: 24832775
- Poschmann G et al. High-fat diet induced isoform changes of the Parkinson's disease protein DJ-1. J Proteome Res 13:2339-51 (2014). PubMed: 24646099
- Besong Agbo D et al. Development of a capillary isoelectric focusing immunoassay to measure DJ-1 isoforms in biological samples. Anal Biochem 443:197-204 (2013). PubMed: 24055619
- O'Connell K & Ohlendieck K Proteomic DIGE analysis of the mitochondria-enriched fraction from aged rat skeletal muscle. Proteomics 9:5509-24 (2009). PubMed: 19834913