Anti-TRIM25/EFP 抗体 [EPR7315] (ab167154)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7315] to TRIM25/EFP
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-TRIM25/EFP antibody [EPR7315]
TRIM25/EFP 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR7315] to TRIM25/EFP -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human TRIM25/EFP aa 100-200. The exact sequence is proprietary.
Database link: Q14258 -
ポジティブ・コントロール
- Recombinant Human TRIM25/EFP protein (ab152814) can be used as a positive control in WB. HeLa and K562 cell lysates; Human breast tissue. Human pancreas tissue. Rat brain lysate. SP2/0 (mouse spleen) 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. Stable for 12 months at -20°C. -
バッファー
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 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
EPR7315 -
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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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
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ab167154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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Flow Cyt (Intra) |
1/60 - 1/90.
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WB | (1) |
1/2000 - 1/50000. Predicted molecular weight: 71 kDa.
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IHC-P |
1/100 - 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 - 1/1000.
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IP |
1/40 - 1/100.
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特記事項 |
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Flow Cyt (Intra)
1/60 - 1/90. |
WB
1/2000 - 1/50000. Predicted molecular weight: 71 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500 - 1/1000. |
IP
1/40 - 1/100. |
ターゲット情報
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機能
Functions as an ubiquitin E3 ligase and as an ISG15 E3 ligase. Involved in innate immune defense against viruses by mediating ubiquitination of DDX58. Mediates 'Lys-63'-linked polyubiquitination of the DDX58 N-terminal CARD-like region which is crucial for triggering the cytosolic signal transduction that leads to the production of interferons in response to viral infection. Promotes ISGylation of 14-3-3 sigma (SFN), an adapter protein implicated in the regulation of a large spectrum signaling pathway. Mediates estrogen action in various target organs. -
組織特異性
Ubiquitous. -
パスウェイ
Protein modification; protein ubiquitination. -
配列類似性
Contains 1 B30.2/SPRY domain.
Contains 1 RING-type zinc finger. -
ドメイン
The RING-type zinc finger is important for ISG15 E3 ligase activity and autoISGylation. AutoISGylation negatively regulates ISG15 E3 ligase activity.
The C-terminal B30.2/SPRY domain interacts with the first N-terminal CARD domain of DDX58. -
翻訳後修飾
Auto-ISGylated. -
細胞内局在
Cytoplasm. Colocalized with DDX58 at cytoplasmic perinuclear bodies. - Information by UniProt
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参照データベース
- Entrez Gene: 7706 Human
- Entrez Gene: 217069 Mouse
- Entrez Gene: 494338 Rat
- Omim: 600453 Human
- SwissProt: Q14258 Human
- SwissProt: Q61510 Mouse
- Unigene: 528952 Human
- Unigene: 248445 Mouse
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別名
- E3 ubiquitin/ISG15 ligase TRIM25 antibody
- EFP antibody
- Estrogen responsive finger protein antibody
see all
画像
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All lanes : Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/10000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TRIM25/EFP knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 71 kDaLanes 1 - 4: Merged signal (red and green). Green - ab167154 observed at 71 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab167154 was shown to specifically react with TRIM25/EFP in wild-type HAP1 cells as signal was lost in TRIM25/EFP knockout cells. Wild-type and TRIM25/EFP knockout samples were subjected to SDS-PAGE. Ab167154 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging. -
All lanes : Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/50000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 71 kDaBlocking and diluting buffer: 5% NFDM/TBST
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ab167154 staining TRIM25/EFP in human pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
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ab167154 staining TRIM25/EFP in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab195889 was used as a tubulin counterstain at a dilution of 1/200 and DAPI was used as a nuclear counterstain.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling TRIM25/EFP with purified ab167154 at 1/90 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/2000 dilution + SP2/0 (mouse spleen) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 71 kDa
Additional bands at: 71 kDa. We are unsure as to the identity of these extra bands.Blocking and diluting buffer: 5% NFDM /TBST
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ab167154 immunoprecipitating TRIM25/EFP. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/1000.
Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab167154 in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate. -
Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/10000 dilution + Rat brain lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 71 kDa
Additional bands at: 71 kDa. We are unsure as to the identity of these extra bands.Blocking and diluting buffer: 5% NFDM/TBST
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All lanes : Anti-TRIM25/EFP antibody [EPR7315] (ab167154) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : K562 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution
Predicted band size: 71 kDa -
Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling TRIM25/EFP with ab167154 at 1/100 dilution.
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Immunofluorescent analysis of HeLa cells labeling TRIM25/EFP with ab167154 at 1/100 dilution.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (26)
ab167154 は 26 報の論文で使用されています。
- D'Amico G et al. ERG activity is regulated by endothelial FAK coupling with TRIM25/USP9x in vascular patterning. Development 149:N/A (2022). PubMed: 35723257
- Booy EP et al. The noncoding RNA BC200 associates with polysomes to positively regulate mRNA translation in tumor cells. J Biol Chem 296:100036 (2021). PubMed: 33410401
- Mei P et al. E3 ligase TRIM25 ubiquitinates RIP3 to inhibit TNF induced cell necrosis. Cell Death Differ 28:2888-2899 (2021). PubMed: 33953350
- Kumar A et al. SARS-CoV-2 Nonstructural Protein 1 Inhibits the Interferon Response by Causing Depletion of Key Host Signaling Factors. J Virol 95:e0026621 (2021). PubMed: 34110264
- Smith EM et al. The mTOR regulated RNA-binding protein LARP1 requires PABPC1 for guided mRNA interaction. Nucleic Acids Res 49:458-478 (2021). PubMed: 33332560