重组Anti-Ndufs4抗体[EP7832] (ab137064)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP7832] to Ndufs4
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
概述
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产品名称
Anti-Ndufs4抗体[EP7832]
参阅全部 Ndufs4 一抗 -
描述
兔单克隆抗体[EP7832] to Ndufs4 -
宿主
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P MouseRatHumanIP RatWB MouseRatHuman -
免疫原
Synthetic peptide within Human Ndufs4 aa 150-250 (C terminal). The exact sequence is proprietary.
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阳性对照
- 293T cell lysates, fetal brain and fetal stomach tissue lysates; Human brain and Human stomach tissues Mouse heat lysate, rat heart lysate, mouse kidney, HeLa.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
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: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP7832 -
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
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab137064 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
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IHC-P |
Mouse
Rat
Human
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IP |
Rat
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WB |
Mouse
Rat
Human
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应用 | Ab评论 | 说明 |
---|---|---|
WB | (1) |
1/500 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 20 kDa).
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IHC-P | (3) |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (2) |
1/500.
For unpurified use at 1/50 - 1/100. |
IP |
1/10 - 1/100.
|
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Flow Cyt |
1/60.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurifed use at 1/100 - 1/1000 dilution.
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说明 |
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WB
1/500 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 20 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/500. For unpurified use at 1/50 - 1/100. |
IP
1/10 - 1/100. |
Flow Cyt
1/60. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurifed use at 1/100 - 1/1000 dilution.
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靶标
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功能
Accessory subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I), that is believed not to be involved in catalysis. Complex I functions in the transfer of electrons from NADH to the respiratory chain. The immediate electron acceptor for the enzyme is believed to be ubiquinone. -
疾病相关
Defects in NDUFS4 are a cause of mitochondrial complex I deficiency (MT-C1D) [MIM:252010]. A disorder of the mitochondrial respiratory chain that causes a wide range of clinical disorders, from lethal neonatal disease to adult-onset neurodegenerative disorders. Phenotypes include macrocephaly with progressive leukodystrophy, non-specific encephalopathy, cardiomyopathy, myopathy, liver disease, Leigh syndrome, Leber hereditary optic neuropathy, and some forms of Parkinson disease. -
序列相似性
Belongs to the complex I NDUFS4 subunit family. -
细胞定位
Mitochondrion inner membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 4724 Human
- Entrez Gene: 17993 Mouse
- Entrez Gene: 499529 Rat
- Omim: 602694 Human
- SwissProt: O43181 Human
- SwissProt: Q9CXZ1 Mouse
- SwissProt: Q5XIF3 Rat
- Unigene: 528222 Human
see all -
别名
- AQDQ antibody
- CI 18 antibody
- CI 18 kDa antibody
see all
图片
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Anti-Ndufs4 antibody [EP7832] (ab137064) at 1/50000 dilution (purified) + Rat heart lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 20 kDaBlocking and diluting buffer: 5% NFDM/TBST
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ab137064 (purified) at 1:30 dilution (2μg) immunoprecipitating Ndufs4 in Rat heart lysate.
Lane 1 (input): Rat heart lysate, 10μg
Lane 2 (+): ab137064 & Rat heart lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab137064 in Rat heart 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. -
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ndufs4 with purified ab137064 at 1:60 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor ® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ndusf5 with Purified ab137064 at 1:500 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab7291 anti-Tubulin (mouse mAb) ab150120 AlexaFluor ® 594 Goat anti-Mouse secondary (1:1000,2 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor ® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs4 antibody [EP7832] (ab137064)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling Ndufs4 with Purified ab137064 at 1:50 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs4 antibody [EP7832] (ab137064)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling Ndufs4 with Purified ab137064 at 1:50 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs4 antibody [EP7832] (ab137064)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue sections labeling Ndufs4 with Purified ab137064 at 1:50 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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All lanes : Anti-Ndufs4 antibody [EP7832] (ab137064) at 1/2000 dilution (purified)
Lane 1 : Human fetal brain lysates
Lane 2 : Mouse heart lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 20 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Ndufs4 knockout HAP1 cell lysate (20 µg)
Lane 3: HEK293 cell lysate (20 µg)
Lane 4: Human fetal heart tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab137064 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.
Unpurified ab137064 was shown to recognize Ndufs4 when Ndufs4 knockout samples were used, along with additional cross-reactive bands. Wild-type and Ndufs4 knockout samples were subjected to SDS-PAGE. ab137064 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10 000 respectively 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 1/10000 dilution for 1 h at room temperature before imaging. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs4 antibody [EP7832] (ab137064)
Immunohistochemical analysis of paraffin-embedded Human brain tissue labelling Ndufs4 with unpurified ab137064 at 1/50 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Overlay histogram showing HepG2 cells stained with unpurified ab137064 (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 (ab137064, 1/1000 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) (0.1μg/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 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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All lanes : Anti-Ndufs4 antibody [EP7832] (ab137064) at 1/1000 dilution (unpurified)
Lane 1 : 293T cell lysate
Lane 2 : Fetal brain tissue lysate
Lane 3 : Fetal kidney tissue lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 20 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ndufs4 antibody [EP7832] (ab137064)
Immunohistochemical analysis of paraffin-embedded Human stomach tissue labelling Ndufs4 with unpurified ab137064 at 1/50 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 (1)
ab137064 被引用在 1 文献中.
- Piekutowska-Abramczuk D et al. NDUFB8 Mutations Cause Mitochondrial Complex I Deficiency in Individuals with Leigh-like Encephalomyopathy. Am J Hum Genet 102:460-467 (2018). PubMed: 29429571