重组Anti-ATP6V0D1/P39抗体[EPR18320-38] (ab202899)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18320-38] to ATP6V0D1/P39
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-ATP6V0D1/P39抗体[EPR18320-38]
参阅全部 ATP6V0D1/P39 一抗 -
描述
兔单克隆抗体[EPR18320-38] to ATP6V0D1/P39 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Human fetal kidney, Human fetal brain, HeLa, MCF-7, A431, C6, PC-12 and PC-12 lysates. Mouse kidney and spleen lysates. Rat spleen lysate. IHC-P: Human breast and cervix carcinoma tissue. Mouse and Rat kidney tissue. ICC/IF: HeLa and MCF-7 cells. IP: MCF-7 whole cell lysate. Flow Cyt (intra): HeLa cells.
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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.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18320-38 -
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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Conjugation kits
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab202899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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Flow Cyt (Intra) |
1/800.
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WB |
1/2000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
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IHC-P |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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IP |
1/50.
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说明 |
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Flow Cyt (Intra)
1/800. |
WB
1/2000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa). |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
IP
1/50. |
靶标
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功能
Subunit of the integral membrane V0 complex of vacuolar ATPase. Vacuolar ATPase is responsible for acidifying a variety of intracellular compartments in eukaryotic cells, thus providing most of the energy required for transport processes in the vacuolar system. May play a role in coupling of proton transport and ATP hydrolysis (By similarity). May play a role in cilium biogenesis through regulation of the transport and the localization of proteins to the cilium. -
组织特异性
Ubiquitous. -
序列相似性
Belongs to the V-ATPase V0D/AC39 subunit family. -
细胞定位
Membrane. Localizes to centrosome and the base of the cilium. - Information by UniProt
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数据库链接
- Entrez Gene: 9114 Human
- Entrez Gene: 11972 Mouse
- Entrez Gene: 291969 Rat
- Omim: 607028 Human
- SwissProt: P61421 Human
- SwissProt: P51863 Mouse
- Unigene: 106876 Human
- Unigene: 17708 Mouse
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别名
- 32 kDa accessory protein antibody
- ATP6D antibody
- ATP6DV antibody
see all
图片
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All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/10000 dilution
Lane 1 : Human fetal kidney
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma)
Lane 3 : MCF-7 (Human breast adenocarcinoma cell line)
Lane 4 : A431 (Human epidermoid carcinoma)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/50000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 1 minuteBlocking/dilution buffer: 5% NFDM/TBST.
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Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/2000 dilution + Human fetal brain at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 5 secondsBlocking/dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/2000 dilution
Lane 1 : Mouse kidney
Lane 2 : Mouse spleen
Lane 3 : Rat spleen
Lane 4 : C6 (Rat glial tumor cells)
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma)
Lane 6 : NIH/3T3 (mouse embryo fibroblast cells)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 5 secondsBlocking/dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Human breast carcinoma tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Mouse kidney tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Rat kidney tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ATP6V0D1/P39 with ab202899 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab202899 at 1/500 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7 (Human breast adenocarcinoma) cells labeling ATP6V0D1/P39 with ab202899 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on MCF-7 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab202899 at 1/500 dilution followed byab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Intracellular Flow Cytometry analysis of HeLa cells labelling ATP6V0D1/P39 with ab202899 at 1/800 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluorr® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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ATP6V0D1/P39 was immunoprecipitated from 1mg of MCF-7 (Human breast adenocarcinoma cell line) whole cell lysate with ab202899 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab202899 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: Input MCF-7 whole cell lysate (10 µg). Lane 2: MCF-7 whole cell lysate following precipitation. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202899 in MCF-7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.10 second exposure.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (4)
ab202899 被引用在 4 文献中.
- Ratto E et al. Direct control of lysosomal catabolic activity by mTORC1 through regulation of V-ATPase assembly. Nat Commun 13:4848 (2022). PubMed: 35977928
- Lee JJ et al. Lysosome-associated membrane protein-2 deficiency increases the risk of reactive oxygen species-induced ferroptosis in retinal pigment epithelial cells. Biochem Biophys Res Commun 521:414-419 (2020). PubMed: 31672277
- Puustinen P et al. DNA-dependent protein kinase regulates lysosomal AMP-dependent protein kinase activation and autophagy. Autophagy 16:1871-1888 (2020). PubMed: 31983282
- Xu Y et al. A Bacterial Effector Reveals the V-ATPase-ATG16L1 Axis that Initiates Xenophagy. Cell 178:552-566.e20 (2019). PubMed: 31327526