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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Ndufs4 antibody [EP7832]
Ndufs4 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP7832] to Ndufs4 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Sheep, Goat, Cat, Dog, Pig, Common marmoset非交差種: Eisenia fetida -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- 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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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
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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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP7832 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab137064の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
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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WB | (1) |
1/500 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 20 kDa).
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IHC-P | (10) |
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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特記事項 |
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Flow Cyt (Intra)
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. |
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. |
ターゲット情報
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機能
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. -
関連疾患
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. -
配列類似性
Belongs to the complex I NDUFS4 subunit family. -
細胞内局在
Mitochondrion inner membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 479335 Dog
- Entrez Gene: 4724 Human
- Entrez Gene: 17993 Mouse
- Entrez Gene: 499529 Rat
- Omim: 602694 Human
- SwissProt: O43181 Human
- SwissProt: Q9CXZ1 Mouse
- SwissProt: Q5XIF3 Rat
see all -
別名
- AQDQ antibody
- CI 18 antibody
- CI 18 kDa antibody
see all
画像
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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. -
Intracellular 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) 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) 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) 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. -
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 withunpurified 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? -
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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (6)
ab137064 は 6 報の論文で使用されています。
- Balderas E et al. Mitochondrial calcium uniporter stabilization preserves energetic homeostasis during Complex I impairment. Nat Commun 13:2769 (2022). PubMed: 35589699
- Creed RB et al. Analysis of hemisphere-dependent effects of unilateral intrastriatal injection of α-synuclein pre-formed fibrils on mitochondrial protein levels, dynamics, and function. Acta Neuropathol Commun 10:78 (2022). PubMed: 35606853
- Weber DD et al. Ketogenic diets slow melanoma growth in vivo regardless of tumor genetics and metabolic plasticity. Cancer Metab 10:12 (2022). PubMed: 35851093
- Thau-Zuchman O et al. A new ketogenic formulation improves functional outcome and reduces tissue loss following traumatic brain injury in adult mice. Theranostics 11:346-360 (2021). PubMed: 33391479
- Jing YY et al. Epigenetic regulation of the Warburg effect by H2B monoubiquitination. Cell Death Differ 27:1660-1676 (2020). PubMed: 31685978
- 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