重组Anti-TFEB抗体[EPR22940-151] (ab270604)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22940-151] to TFEB
- Suitable for: ICC/IF, IP, Flow Cyt (Intra), IHC-P, WB
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-TFEB抗体[EPR22940-151]
参阅全部 TFEB 一抗 -
描述
兔单克隆抗体[EPR22940-151] to TFEB -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, IP, Flow Cyt (Intra), IHC-P, WBmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: IM-9 and Raji whole cell lysate. IHC-P: Human lung cancer and colon tissue. ICC/IF: IM-9 and Raji cells. Flow Cyt (intra): IM-9 and Raji cells. IP: Raji whole cell lysate.
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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: 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
EPR22940-151 -
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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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab270604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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ICC/IF |
1/50.
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IP |
1/30.
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Flow Cyt (Intra) |
1/500.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Detects a band of approximately 66 kDa (predicted molecular weight: 53 kDa).
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说明 |
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ICC/IF
1/50. |
IP
1/30. |
Flow Cyt (Intra)
1/500. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 66 kDa (predicted molecular weight: 53 kDa). |
靶标
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功能
Transcription factor that specifically recognizes and binds E-box sequences (3'-CANNTG-5'). Efficient DNA-binding requires dimerization with itself or with another MiT/TFE family member such as TFE3 or MITF. In association with TFE3, activates the expression of CD40L in T-cells, thereby playing a role in T-cell-dependent antibody responses in activated CD4(+) T-cells and thymus-dependent humoral immunity. Specifically recognizes and binds the CLEAR-box sequence (5'-GTCACGTGAC-3') present in the regulatory region of many lysosomal genes, leading to activate their expression. It thereby plays a central role in expression of lysosomal genes. Specifically recognizes the gamma-E3 box, a subset of E-boxes, present in the heavy-chain immunoglobulin enhancer. Plays a role in the signal transduction processes required for normal vascularization of the placenta. -
序列相似性
Belongs to the MiT/TFE family.
Contains 1 basic helix-loop-helix (bHLH) domain. -
结构域
The leucin zipper region is essential for homo- or heterodimerization and high-affinity DNA binding. DNA binding is mediated by the basic region. -
翻译后修饰
Sumoylated; does not affect dimerization with MITF. -
细胞定位
Cytoplasm. Nucleus. Mainly present in the cytoplasm. Under aberrant lysosomal storage conditions, it translocates from the cytoplasm to the nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 7942 Human
- Omim: 600744 Human
- SwissProt: P19484 Human
- Unigene: 485360 Human
- Unigene: 715310 Human
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别名
- Alpha TFEB antibody
- AlphaTFEB antibody
- bHLHe35 antibody
see all
图片
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Flow cytometry overlay histogram showing left Raji positive cells and right negative MCF7 stained with ab270604 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab270604) (1x 106 in 100μl at 0.2μg/ml (1/10350)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-TFEB antibody [EPR22940-151] (ab270604) at 1/1000 dilution
Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 2 : IM-9 (human multiple myeloma B Lymphoblast), whole cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 66 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:30519051).
Exposure time: 26 secs.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling TFEB with ab270604 at 1/500 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human colon tissue is observed (PMID:30519051). The section was incubated with ab270604 for 20 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji cells labeling TFEB with ab270604 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Raji cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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TFEB was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate with ab270604 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270604 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.
Lane 1: Raji whole cell lysate.
Lane 2: ab270604 IP in Raji whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270604 in Raji whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 secs.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized IM-9 (Human multiple myeloma B Lymphoblast) cells labeling TFEB with ab270604 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling TFEB with ab270604 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized IM-9 cells labeling TFEB with ab270604 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in IM-9 cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TFEB with ab270604 at 1/500 followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer tissue is observed (PMID:26264650). The section was incubated with ab270604 for 20 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (9)
ab270604 被引用在 9 文献中.
- Mao X et al. Lipid reprogramming induced by the TFEB-ERRα axis enhanced membrane fluidity to promote EC progression. J Exp Clin Cancer Res 41:28 (2022). PubMed: 35045880
- Liu Y et al. Toll-Like Receptor 4 Exacerbates Mycoplasma pneumoniaevia Promoting Transcription Factor EB-Mediated Autophagy. Contrast Media Mol Imaging 2022:3357694 (2022). PubMed: 35965629
- Fan L et al. Melatonin Ameliorates the Progression of Alzheimer's Disease by Inducing TFEB Nuclear Translocation, Promoting Mitophagy, and Regulating NLRP3 Inflammasome Activity. Biomed Res Int 2022:8099459 (2022). PubMed: 35983247
- Mahali S et al. Defective proteostasis in induced pluripotent stem cell models of frontotemporal lobar degeneration. Transl Psychiatry 12:508 (2022). PubMed: 36494352
- Zhou W et al. Enhanced Autolysosomal Function Ameliorates the Inflammatory Response Mediated by the NLRP3 Inflammasome in Alzheimer's Disease. Front Aging Neurosci 13:629891 (2021). PubMed: 33708103
- Xu L & Pan J Transcription factor EB promotes rheumatoid arthritis of Sprague-Dawley rats via regulating autophagy. 3 Biotech 11:162 (2021). PubMed: 33786279
- Xiong Y et al. Virucidal activity of Moringa A from Moringa oleifera seeds against Influenza A Viruses by regulating TFEB. Int Immunopharmacol 95:107561 (2021). PubMed: 33744778
- Wang L et al. Torin 1 alleviates impairment of TFEB-mediated lysosomal biogenesis and autophagy in TGFBI (p.G623_H626del)-linked Thiel-Behnke corneal dystrophy. Autophagy N/A:1-18 (2021). PubMed: 34403298
- Zhan HQ et al. TFEB promotes BCL-2 expression by upregulating its promoter activity in the t(6;11) translocation renal cell carcinomas. Am J Transl Res 13:8804-8818 (2021). PubMed: 34539996