Anti-ATP5A抗体[15H4C4] - Mitochondrial Marker (ab14748)
Key features and details
- Mouse monoclonal [15H4C4] to ATP5A - Mitochondrial Marker
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
- Reacts with: Mouse, Rat, Cow, Human, Drosophila melanogaster
- Isotype: IgG2b
Related conjugates and formulations
概述
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产品名称
Anti-ATP5A抗体[15H4C4] - Mitochondrial Marker
参阅全部 ATP5A 一抗 -
描述
小鼠单克隆抗体[15H4C4] to ATP5A - Mitochondrial Marker -
宿主
Mouse -
经测试应用
适用于: WB, IHC-P, ICC/IF, Flow Cytmore details -
种属反应性
与反应: Mouse, Rat, Cow, Human, Drosophila melanogaster
预测可用于: Pig -
免疫原
Tissue, cells or virus. This information is considered to be commercially sensitive.
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阳性对照
- WB: Isolated mitochondria from human, cow, rat and mouse heart. Human liver tissue lysate. HepG2 whole cell lysate. ICC/IF: HeLa, MCF7 and MDA-MB-231 cells. IHC-P: Human heart tissue. Flow Cyt: HepG2 cells.
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常规说明
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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. Store at +4°C. -
存储溶液
pH: 7.5
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading...
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纯度
IgG fraction -
纯化说明
Near homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. -
克隆
单克隆 -
克隆编号
15H4C4 -
同种型
IgG2b -
轻链类型
kappa -
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ab14748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 | (12) |
Use a concentration of 1 µg/ml. Detects a band of approximately 53 kDa.
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IHC-P | (6) |
Use a concentration of 1 µg/ml.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF | (3) |
Use a concentration of 1 - 10 µg/ml.
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Flow Cyt |
Use a concentration of 1 µg/ml.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
说明 |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 53 kDa. |
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 1 - 10 µg/ml. |
Flow Cyt
Use a concentration of 1 µg/ml. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Mitochondrial membrane ATP synthase (F(1)F(0) ATP synthase or Complex V) produces ATP from ADP in the presence of a proton gradient across the membrane which is generated by electron transport complexes of the respiratory chain. F-type ATPases consist of two structural domains, F(1) - containing the extramembraneous catalytic core, and F(0) - containing the membrane proton channel, linked together by a central stalk and a peripheral stalk. During catalysis, ATP synthesis in the catalytic domain of F(1) is coupled via a rotary mechanism of the central stalk subunits to proton translocation. Subunits alpha and beta form the catalytic core in F(1). Rotation of the central stalk against the surrounding alpha(3)beta(3) subunits leads to hydrolysis of ATP in three separate catalytic sites on the beta subunits. Subunit alpha does not bear the catalytic high-affinity ATP-binding sites. -
组织特异性
Fetal lung, heart, liver, gut and kidney. Expressed at higher levels in the fetal brain, retina and spinal cord. -
序列相似性
Belongs to the ATPase alpha/beta chains family. -
翻译后修饰
The N-terminus is blocked. -
细胞定位
Mitochondrion inner membrane. Peripheral membrane protein. - Information by UniProt
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数据库链接
- Entrez Gene: 282578 Cow
- Entrez Gene: 37617 Drosophila melanogaster
- Entrez Gene: 498 Human
- Entrez Gene: 11946 Mouse
- Entrez Gene: 100157880 Pig
- Entrez Gene: 65262 Rat
- Omim: 164360 Human
- SwissProt: P19483 Cow
see all -
别名
- ATP synthase alpha chain antibody
- ATP synthase alpha chain, mitochondrial antibody
- ATP synthase subunit alpha antibody
see all
图片
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Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling ATP5A with ab14748 at a concentration of 0.01µg/ml.
The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5. ab14748 anti-ATP5A was incubated at 37°C for 16min.
Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
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Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling ATP5A with ab14748 at a concentration of 0.05µg/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.
ab14748 anti-ATP5A antibody [15H4C4] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
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Testes of bb8ms mutants show defects in post-meiotic, elongated spermatids.
(A-B) Spermatids from WT (A) and bb8ms (B) testis both have elongated cysts, but there are large spherical vesicles in the mutant (arrows) by phase contrast microscopy. Scale bars: 100 μm.
(C, D) Mitochondria of elongated spermatids stained with ATP5α antibody ab14748 in WT (C) and in bb8ms (D) mutants. ATP5α positive staining of the large vesicles in the cysts are indicated by arrow. Scale bars: 50 μm.
(E, F) JC-1 staining positive large vesicles (arrows) are absent from WT (E), but present in bb8ms elongated cysts (F). Scale bars: 25 μm.
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Western blots were probed for HNE-damaged mitochondrial protein from brainstem (A), cerebellum (B) and “rest” brain (R). The sample designation indicates the age group (y for P25-P35, o for P45-P55), the genotype (KO, Ctrl for controls) and a number to distinguish independent samples.
Panel D is the blot from panel A reprobed for the mitochondrial marker ATPase (ATP5a) using ab14748 to demonstrate that extended sample storage did not degrade sample protein in general. Black lines in the MW lanes are magic marker on the film to indicate the positions of the prestained molecular weight standards on the blot.
For full image please see paper.
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All lanes : Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (ab14748) at 1 µg/ml
Lane 1 : Isolated mitochondria from human heart at 10 µg
Lane 2 : Isolated mitochondria from bovine heart at 4 µg
Lane 3 : Isolated mitochondria from rat heart at 10 µg
Lane 4 : Isolated mitochondria from mouse heart at 10 µg
Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) lysate at 20 µg -
All lanes : Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (ab14748) at 1 µg/ml
Lane 1 : Human liver tissue lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Observed band size: 55 kDa why is the actual band size different from the predicted?
Additional bands at: 36 kDa. We are unsure as to the identity of these extra bands. -
ICC/IF image of ab14748 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in 100% methanol (5 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab14748 at 1 µg/ml (shown in red) and ab6160 (Rat monoclonal to Tubulin) at 1 µg/ml (shown in green).
This was followed by an incubation at room temperature for 1 hour with ab150119, Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed, at 0.5 µg/ml (shown in red) and ab150165, Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed, at 0.5 µg/ml (shown in green).
Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 minutes).
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ab14748 staining ATP5A in MDA-MB-231 cells by ICC/IF (Immunocytochemistry/immunofluorescence).
Cells were fixed with formaldehyde, permeabilized with 1% Triton X-100 and blocked with 10% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/100 in BSA + 0.02% Tween 20) for 1 hour at 21°C. A DyLight® 550-conjugated goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.
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ICC/IF image of ab14748 stained MCF7 (Human breast adenocarcinoma cell line) cells.
The cells were fixed in 4% formaldehyde (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14748, 10 µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
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ab14748 (1 µg/ml) staining ATP5A in human heart (left ventricle), using an automated system (DAKO Autostainer Plus). Using this protocol there is mitochondrial staining of cardiomyocytes.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes.Slides were counterstained with hematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
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Overlay histogram showing HepG2 (Human liver hepatocellular carcinoma cell line) cells stained with ab14748 (red line).
The cells were fixed with 4% paraformaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab14748, 1 µg/1x106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2 µg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events was performed.
This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.
数据表及文件
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SDS download
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Datasheet download
文献 (553)
ab14748 被引用在 553 文献中.
- Moschandrea C et al. Mitochondrial dysfunction abrogates dietary lipid processing in enterocytes. Nature 625:385-392 (2024). PubMed: 38123683
- Gorbacheva EY et al. The Oxidative Phosphorylation and Cytoskeleton Proteins of Mouse Ovaries after 96 Hours of Hindlimb Suspension. Life (Basel) 13:N/A (2023). PubMed: 38137934
- Burgin H et al. Loss of mitochondrial fatty acid β-oxidation protein short-chain Enoyl-CoA hydratase disrupts oxidative phosphorylation protein complex stability and function. FEBS J 290:225-246 (2023). PubMed: 35962613
- Lin G et al. Exploring therapeutic strategies for infantile neuronal axonal dystrophy (INAD/PARK14). Elife 12:N/A (2023). PubMed: 36645408
- Wang B et al. The mitochondrial Ahi1/GR participates the regulation on mtDNA copy numbers and brain ATP levels and modulates depressive behaviors in mice. Cell Commun Signal 21:21 (2023). PubMed: 36691038