重组Anti-PARK7/DJ1抗体[EP2815Y] - BSA and Azide free (ab218373)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2815Y] to PARK7/DJ1 - BSA and Azide free
- Suitable for: IP, ICC/IF, WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PARK7/DJ1抗体[EP2815Y] - BSA and Azide free
参阅全部 PARK7/DJ1 一抗 -
描述
兔单克隆抗体[EP2815Y] to PARK7/DJ1 - BSA and Azide free -
宿主
Rabbit -
特异性
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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经测试应用
适用于: IP, ICC/IF, WB, IHC-P, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Jurkat, HeLa, NIH3T3 or 293T cell lysate. Human fetal brain; Human brain nuclear fraction tissue lysate; Mouse brain and Rat brain tissue lysates. IHC-P: Human Lung and Brain tissue. ICC/IF: PANC-1 and Jurkat cell lines. Flow Cyt (intra): HepG2 cells. IP: Mouse brain lysate.
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常规说明
ab218373 is the carrier-free version of ab76008.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP2815Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-PARK7/DJ1 antibody [EP2815Y] (ab203989)
- Alexa Fluor® 647 Anti-PARK7/DJ1 antibody [EP2815Y] (ab204009)
- Alexa Fluor® 594 Anti-PARK7/DJ1 antibody [EP2815Y] (ab215102)
- PE Anti-PARK7/DJ1 antibody [EP2815Y] (ab305853)
- APC Anti-PARK7/DJ1 antibody [EP2815Y] (ab305854)
- HRP Anti-PARK7/DJ1 antibody [EP2815Y] (ab305855)
- Alexa Fluor® 555 Anti-PARK7/DJ1 antibody [EP2815Y] (ab307128)
- Anti-PARK7/DJ1 antibody [EP2815Y] (ab76008)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab218373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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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 20 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Perform heat mediated antigen retrieval using 0.01M Sodium Citrate Buffer, pH 6.0 before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376- Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
说明 |
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IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 20 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Perform heat mediated antigen retrieval using 0.01M Sodium Citrate Buffer, pH 6.0 before commencing with IHC staining protocol. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376- Rabbit monoclonal IgG (Low endotoxin, Azide free), 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 [EP2815Y] (ab76008) 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?This data was developed using ab76008, the same antibody clone in a different buffer formulation.
Lanes 1-4: Merged signal (red and green). Green - ab76008 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
ab76008 Anti-PARK7/DJ1 antibody [EP2815Y] 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. ab76008 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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This IHC data was generated using the same anti-PARK7 antibody clone, EP2815Y, in a different buffer formulation (cat# ab76008).
Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling PARK7/DJ1 with Purified ab76008 at 1:500 dilution (0.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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This IHC data was generated using the same anti-PARK7 antibody clone, EP2815Y, in a different buffer formulation (cat# ab76008).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling PARK7/DJ1 with Purified ab76008 at 1:1000 dilution (0.11 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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This IHC data was generated using the same anti-PARK7 antibody clone, EP2815Y, in a different buffer formulation (cat
ab76008).
Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling PARK7/DJ1 with Purified ab76008 at 1/20 dilution (10µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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This WB data was generated using the same anti-PARK7 antibody clone, EP2815Y, in a different buffer formulation (cat# ab76008).
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PARK7/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 - ab76008 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.ab76008 was shown to specifically react with PARK/DJ1 when PARK/DJ1 knockout samples were used. Wild-type and PARK/DJ1 knockout samples were subjected to SDS-PAGE. ab76008 and ab8245 (loading control to GAPDH) 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 1 h at room temperature before imaging.
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This IHC data was generated using the same anti-PARK7 antibody clone, EP2815Y, in a different buffer formulation (cat# ab76008).
ab76008 (purified) at 1:20 dilution (0.5µg) immunoprecipitating PARK7/DJ1 in Mouse brain lysate.
Lane 1 (input): Mouse brain lysate 10µg
Lane 2 (+): ab76008 & Mouse brain lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76008 in Mouse brain lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling PARK7/DJ1 with purified ab76008 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76008).
This image was generated using the unpurified version of the product.
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Overlay histogram showing HepG2 cells stained with ab76008 (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 (ab76008, 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 HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76008).
This image was generated using the unpurified version of the product.
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This IHC data was generated using the same anti-PARK7 antibody clone, EP2815Y, in a different buffer formulation (cat# ab76008).
ab76008, at 1/250 dilution, staining PARK7/DJ1 in human brain by immunohistochemistry using paraffin-embedded tissue.
This image was generated using the unpurified version of the product.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (6)
ab218373 被引用在 6 文献中.
- Vázquez-Mayorga E et al. Novel Redox-Dependent Esterase Activity (EC 3.1.1.2) for DJ-1: Implications for Parkinson's Disease. Int J Mol Sci 17:N/A (2016). PubMed: 27556455
- Kudinov AE et al. Musashi-2 (MSI2) supports TGF-ß signaling and inhibits claudins to promote non-small cell lung cancer (NSCLC) metastasis. Proc Natl Acad Sci U S A 113:6955-60 (2016). PubMed: 27274057
- Wang B et al. Essential control of mitochondrial morphology and function by chaperone-mediated autophagy through degradation of PARK7. Autophagy 12:1215-28 (2016). PubMed: 27171370
- Moscovitz O et al. The Parkinson's-associated protein DJ-1 regulates the 20S proteasome. Nat Commun 6:6609 (2015). WB . PubMed: 25833141
- Choi HK et al. PINK1 positively regulates HDAC3 to suppress dopaminergic neuronal cell death. Hum Mol Genet 24:1127-41 (2015). PubMed: 25305081
- Kaneko Y et al. DJ-1 ameliorates ischemic cell death in vitro possibly via mitochondrial pathway. Neurobiol Dis 62:56-61 (2014). ICC ; Human . PubMed: 24060818