重组Anti-FTO抗体[EPR6894] (ab126605)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6894] to FTO
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-FTO抗体[EPR6894]
参阅全部 FTO 一抗 -
描述
兔单克隆抗体[EPR6894] to FTO -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IFmore details
不适用于: Flow Cyt -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human FTO (N terminal). The exact sequence is proprietary.
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阳性对照
- WB: Wild-type MCF7, HEK-293, MOLT-4, BxPC-3, Caco-2, SH-SY5Y and 293T cell lysates. IHC-P: Human hepatocellular carcinoma and human adrenal gland tissues. ICC/IF: BxPC-3 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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 59% PBS, 40% Glycerol -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR6894 -
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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KO cell lines
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab126605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 |
1/10000 - 1/20000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
For unpurified, use 1/1000 - 1/10000. |
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IHC-P | (1) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
For unpurified, use 1/50 - 1/100. |
ICC/IF | (1) |
1/250.
For unpurified, use 1/50 - 1/100. |
说明 |
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WB
1/10000 - 1/20000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa). For unpurified, use 1/1000 - 1/10000. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. For unpurified, use 1/50 - 1/100. |
ICC/IF
1/250. For unpurified, use 1/50 - 1/100. |
靶标
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功能
Dioxygenase that repairs alkylated DNA and RNA by oxidative demethylation. Has highest activity towards single-stranded RNA containing 3-methyluracil, followed by single-stranded DNA containing 3-methylthymine. Has low demethylase activity towards single-stranded DNA containing 1-methyladenine or 3-methylcytosine. Has no activity towards 1-methylguanine. Has no detectable activity towards double-stranded DNA. Requires molecular oxygen, alpha-ketoglutarate and iron. Contributes to the regulation of the global metabolic rate, energy expenditure and energy homeostasis. Contributes to the regulation of body size and body fat accumulation. -
组织特异性
Ubiquitously expressed, with relatively high expression in adrenal glands and brain; especially in hypothalamus and pituitary. -
疾病相关
Defects in FTO are the cause of growth retardation developmental delay coarse facies and early death (GRDDCFED) [MIM:612938]. The disease consists of a severe children multiple congenital anomaly syndrome with death by the age of 3 years. All affected individuals had postnatal growth retardation, microcephaly, severe psychomotor delay, functional brain deficits, and characteristic facial dysmorphism. In some patients, structural brain malformations, cardiac defects, genital anomalies, and cleft palate were also observed. -
序列相似性
Belongs to the fto family. -
结构域
The 3D-structure of the Fe2OG dioxygenase domain is similar to that of the Fe2OG dioxygenase domain found in the bacterial DNA repair dioxygenase alkB and its mammalian orthologs, but sequence similarity is very low. As a consequence, the domain is not detected by protein signature databases. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 79068 Human
- Omim: 610966 Human
- SwissProt: Q9C0B1 Human
- Unigene: 528833 Human
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别名
- AlkB homolog 9 antibody
- ALKBH9 antibody
- Alpha-ketoglutarate-dependent dioxygenase FTO antibody
see all
图片
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All lanes : Anti-FTO antibody [EPR6894] (ab126605) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : FTO knockout MCF7 cell lysate
Lane 3 : HEK-293 cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 58 kDaFalse colour image of Western blot: Anti-FTO antibody [EPR6894] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab126605 was shown to bind specifically to FTO. A band was observed at 58 kDa in wild-type MCF7 cell lysates with no signal observed at this size in FTO knockout cell line. To generate this image, wild-type and FTO knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: FTO knockout HAP1 whole cell lysate (20 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: MOLT4 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab126605 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab126605 was shown to specifically react with FTO in wild-type HAP1 cells. No band was observed when FTO knockout samples were examined. Wild-type and FTO knockout samples were subjected to SDS-PAGE. ab126605 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical staining of paraffin embedded human hepatocellular carcinoma with purified ab126605 at a working dilution of 1 in 500. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunofluorescence staining of BxPC-3 cells with purified ab126605 at a working dilution of 1 in 250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab126605 was used at a dilution of 1/200 followed by an Alexa Fluor® 488 goat anti-mouse antibody at a dilution of 1/500.
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All lanes : Anti-FTO antibody [EPR6894] (ab126605) at 1/10000 dilution (purified)
Lane 1 : Molt-4 cell lysate
Lane 2 : HEK293 cell lysate
Lane 3 : BxPC-3 cell lysate
Lane 4 : Caco-2 cell lysate
Lane 5 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 58 kDa
Observed band size: 58 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Immunohistochemical staining of FTO in paraffin embedded human adrenal gland tissue using unpurified ab126605 at a 1/50 dilution.
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Unpurified ab126605 at a 1/50 dilution staining FTO in BxPC3 cells by immunofluorescence.
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All lanes : Anti-FTO antibody [EPR6894] (ab126605) at 1/1000 dilution (unpurified)
Lane 1 : MOLT4 cell lysate
Lane 2 : 293T cell lysate
Lane 3 : SH SY5Y cell lysate
Lane 4 : BxPC3 cell lysate
Lane 5 : Caco 2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 58 kDa
Observed band size: 58 kDa
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (28)
ab126605 被引用在 28 文献中.
- Liu X et al. m6A methylated EphA2 and VEGFA through IGF2BP2/3 regulation promotes vasculogenic mimicry in colorectal cancer via PI3K/AKT and ERK1/2 signaling. Cell Death Dis 13:483 (2022). WB ; Human . PubMed: 35595748
- Wang R et al. FTO protects human granulosa cells from chemotherapy-induced cytotoxicity. Reprod Biol Endocrinol 20:39 (2022). PubMed: 35219326
- Zhang S et al. SPI1-induced downregulation of FTO promotes GBM progression by regulating pri-miR-10a processing in an m6A-dependent manner. Mol Ther Nucleic Acids 27:699-717 (2022). PubMed: 35317283
- Ji FH et al. FTO Prevents Thyroid Cancer Progression by SLC7A11 m6A Methylation in a Ferroptosis-Dependent Manner. Front Endocrinol (Lausanne) 13:857765 (2022). PubMed: 35721711
- Zhang Y et al. M6 A demethylase fat mass and obesity-associated protein regulates cisplatin resistance of gastric cancer by modulating autophagy activation through ULK1. Cancer Sci 113:3085-3096 (2022). PubMed: 35730319