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
製品の概要
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製品名
Anti-TFEB antibody [EPR22940-151]
TFEB 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR22940-151] to TFEB -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, IP, Flow Cyt (Intra), IHC-P, WBmore details -
種交差性
交差種: Human -
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
- 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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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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
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
EPR22940-151 -
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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Related Products
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ab270604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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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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特記事項 |
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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). |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the MiT/TFE family.
Contains 1 basic helix-loop-helix (bHLH) domain. -
ドメイン
The leucin zipper region is essential for homo- or heterodimerization and high-affinity DNA binding. DNA binding is mediated by the basic region. -
翻訳後修飾
Sumoylated; does not affect dimerization with MITF. -
細胞内局在
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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参照データベース
- Entrez Gene: 7942 Human
- Omim: 600744 Human
- SwissProt: P19484 Human
- Unigene: 485360 Human
- Unigene: 715310 Human
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別名
- Alpha TFEB antibody
- AlphaTFEB antibody
- bHLHe35 antibody
see all
画像
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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.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (9)
ab270604 は 9 報の論文で使用されています。
- 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