Anti-Pan Trk 抗体 [EP1058Y] (ab76291)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1058Y] to Pan Trk
- Suitable for: Indirect ELISA, ICC/IF, Flow Cyt (Intra), WB, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Pan Trk antibody [EP1058Y]
Pan Trk 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1058Y] to Pan Trk -
由来種
Rabbit -
特異性
This antibody detects both phosphorylated and unphosphorylated Pan Trk. Based on the WB and FC data, this antibody has relatively lower affinity to TrkC compared to TrkA and TrkB.
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アプリケーション
適用あり: Indirect ELISA, ICC/IF, Flow Cyt (Intra), WB, IP, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, 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: Human, mouse and rat brain tissue lysates IHC-P: Human, mouse and rat cerebrum tissue, His-human TrkA/B and C overexpression 293T whole cell pellet. ICC/IF: U87-MG and SH-SY5Y cells; Mouse DRG neurons; mouse primary neuron. Flow Cyt (intra): SH-SY5Y cells. IP: Rat and mouse brain tissue 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. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% 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
EP1058Y -
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
- Rat brain normal tissue lysate - membrane extract (ab29473)
- Mouse brain tissue lysate - total protein (ab30151)
- Rat brain cerebellum tissue lysate - total protein (ab4032)
- Mouse brain tissue lysate - total protein (0 days) (ab7188)
- Mouse brain tissue lysate - total protein (14 days) (ab7189)
- Mouse brain tissue lysate - total protein (7 days) (ab7190)
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ab76291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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Indirect ELISA |
Use at an assay dependent concentration.
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ICC/IF | (1) |
1/100.
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Flow Cyt (Intra) |
1/130.
Paraformaldehyde or methanol fixed cells, permeabilization recommended. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (2) |
1/1000. Predicted molecular weight: 87 kDa.
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IP |
1/40.
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IHC-P | (3) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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特記事項 |
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Indirect ELISA
Use at an assay dependent concentration. |
ICC/IF
1/100. |
Flow Cyt (Intra)
1/130. Paraformaldehyde or methanol fixed cells, permeabilization recommended. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000. Predicted molecular weight: 87 kDa. |
IP
1/40. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ターゲット情報
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関連性
Family of neurotrophic tyrosine kinase (NTRK1/2/3) genes which encode TrkA, TrkB and TrkC protein kinases. The three family members are activated by different neurotrophins: TrkA is activated by Nerve growth factor (NGF), TrkB by Brain-derived neurotrophic factor (BDNF) or neurotrophin-4 (NT-4) and TrkC by NT-3. Neurotrophin signalling activates cellular pathways involved in the development and the maturation of the central and peripheral nervous systems through regulation of proliferation, differentiation and survival of sympathetic and nervous neurons. Localization TrkA: Cell membrane. Early endosome membrane. Late endosome membrane. Internalized to endosomes upon binding of NGF or NT-3 and further transported to the cell body via a retrograde axonal transport. Localized at cell membrane and early endosomes before nerve growth factor (NGF) stimulation. Recruited to late endosomes after NGF stimulation. Colocalized with RAPGEF2 at late endosomes (By similarity). TrkB: Membrane. TrkC: Membrane. -
参照データベース
- Entrez Gene: 4914 Human
- Entrez Gene: 4915 Human
- Entrez Gene: 4916 Human
- Entrez Gene: 18211 Mouse
- Entrez Gene: 18212 Mouse
- Entrez Gene: 18213 Mouse
- Entrez Gene: 25054 Rat
- Entrez Gene: 29613 Rat
see all -
別名
- BDNF/NT-3 growth factors receptor antibody
- EML4 antibody
- ETV6 antibody
see all
画像
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of (A) His-human TrkA overexpression 293T whole cell pellet, (B) His-human TrkB overexpression 293T whole cell pellet, (C) His-human TrkC overexpression 293T whole cell pellet and (D) HEK-293T transfected with empty plasmid labelling Pan Trk with purified ab76291 at 1/1000. Heat mediated antigen retrieval was performed using Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes. Sections were counterstained with hematoxylin.
Positive staining on (A) His-human TrkA overexpression 293T whole cell pellet, (B) His-human TrkB overexpression 293T whole cell pellet and (C) His-human TrkC overexpression 293T whole cell pellet.
No staining on (D) HEK-293T transfected with empty plasmid.
The section was incubated with abab76291 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument -
All lanes : Anti-Pan Trk antibody [EP1058Y] (ab76291) at 1/1000 dilution
Lanes 1 & 3 & 5 : Empty vector over expression 293T whole cell lysates
Lane 2 : His-human TrkA overexpression 293T whole cell lysates
Lane 4 : His-human TrkB overexpression 293T whole cell lysates
Lane 6 : His-human TrkC overexpression 293T whole cell lysates
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: 87 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST
This antibody has relatively lower affinity to TrkC compared to TrkA and TrkB.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue labelling Pan Trk with purified ab76291 at 1/1000. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Heat mediated antigen retrieval was performed using Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes. Sections were counterstained with hematoxylin. Negative control using PBS instead of primary antibody.
Positive staining on mouse cerebrum.
The section was incubated with ab76291 for 30 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. -
Immunocytochemistry/immunofluorescence analysis of SH-SY5Y (Human neuroblastoma epithelial cell) labeling pan Trk with ab76291 at 1/100 dilution (7 μg/mL). ab150077, AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/mL) was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. DAPI (blue) was used as nuclear counterstain. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 μg/mL).
Confocal image showing cytoplasmic staining in SH-SY5Y cell line.
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All lanes : Anti-Pan Trk antibody [EP1058Y] (ab76291) at 1/1000 dilution
Lane 1 : Human brain lysates
Lane 2 : Human kidney lysates
Lane 3 : Mouse brain lysates
Lane 4 : Mouse kidney lysates
Lane 5 : Rat brain lysates
Lane 6 : Rat kidney lysates
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: 87 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?
Exposure time: 4 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling Pan Trk with ab76291 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. 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).
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Intracellular Flow Cytometry analysis of293T (human embryonic kidney epithelial cell) transfected with TrkB overexpression construct labeling Pan Trk with ab76291 at 1/700 dilution (0.1 µg) (Right). The cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor®488, ab150077) at 1/2000 dilution was used as the secondary antibody.Isotype control/colour: Rabbit monoclonal IgG (ab172730) (Left).
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Intracellular Flow Cytometry analysis of293T (human embryonic kidney epithelial cell) transfected with TrkA overexpression construct labeling Pan Trk with ab76291 at 1/700 dilution (0.1 µg) (Right). The cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.Isotype control/colour: Rabbit monoclonal IgG (ab172730) (Left).
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Immunocytochemistry/immunofluorescence analysis of TrkA- overexpressed 293T (human embryonic kidney epithelial cell) labelled with the pan Trk antibody ab76291 at 1/1000 dilution (0.7 μg/mL). ab150077, AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/mL) was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. DAPI (blue) was used as nuclear counterstain. Cells were counterstained with Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 647 Conjugate) at 1/200 (2.5 μg/mL).
Confocal image showing cytoplasmic staining in 293T cells transfected with a myc-tagged hTrkA expression vector.
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Immunocytochemistry/immunofluorescence analysis of TrkB- overexpressed 293T (human embryonic kidney epithelial cell) labelled with the pan Trk antibody ab76291 at 1/1000 dilution (0.7 μg/mL). ab150077, AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/mL) was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. DAPI (blue) was used as nuclear counterstain. Cells were counterstained with Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 647 Conjugate) at 1/200 (2.5 μg/mL).
Confocal image showing cytoplasmic staining in 293T cells transfected with a myc-tagged hTrkB expression vector.
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Immunocytochemistry/immunofluorescence analysis of TrkC- overexpressed 293T (human embryonic kidney epithelial cell) labelled with the pan Trk antibody ab76291 at 1/1000 dilution (0.7 μg/mL). ab150077, AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/mL) was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. DAPI (blue) was used as nuclear counterstain. Cells were counterstained with Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 647 Conjugate) at 1/200 (2.5 μg/mL).
Confocal image showing cytoplasmic staining in 293T cells transfected with a myc-tagged hTrkC expression vector.
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ELISA analysis of Human TrkA recombinant protein at 250 ng/mL with ab76291. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
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ab76291 at 1/40 immunoprecipitating Pan Trk in human brain tissue lysate observed at 145 kDa.
Lane 1 (input): Human brain tissue lysate (10µg)
Lane 2 (+): ab76291+ human brain tissue lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76291 in human brain lysate
For western blotting, ab76291 at 1/1000 dilution (0.7 μg/mL) and VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 were used.The 30 kDa band is an intracellular fragment, and the 140 kDa observed MW which is higher than the predicted one is due to the glycosylation modification. (refer to ab189903).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
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ab76291 at 1/40 immunoprecipitating Pan Trk in mouse brain tissue lysate observed at 145 kDa.
Lane 1 (input): Mouse brain tissue lysate (10µg)
Lane 2 (+): ab76291+ mouse brain tissue lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76291 in Mouse brain lysate
For western blotting, ab76291 at 1/1000 dilution (0.7 μg/mL) and VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 were used.The 30 kDa band is an intracellular fragment, and the 140 kDa observed MW which is higher than the predicted one is due to the glycosylation modification. (refer to ab189903).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
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Overlay histogram showing SH-SY5Y (human neuroblastoma cell line from bone marrow) cells stained with unpurified ab76291 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76291, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with methanol (5 min) used under the same conditions.Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue labelling Pan Trk with purified ab76291 at 1/1000. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Heat mediated antigen retrieval was performed using Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes. Sections were counterstained with hematoxylin. Negative control using PBS instead of primary antibody.
Positive staining on human cerebrum.
The section was incubated with ab76291 for 30 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue labelling Pan Trk with purified ab76291 at 1/1000. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Heat mediated antigen retrieval was performed using Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes. Sections were counterstained with hematoxylin. Negative control using PBS instead of primary antibody.
Positive staining on rat cerebrum.
The section was incubated with ab76291 for 30 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. -
All lanes : Anti-Pan Trk antibody [EP1058Y] (ab76291) at 1/5000 dilution (unpurified)
Lane 1 : Rat brain tissue lysate (untreated)
Lane 2 : Rat brain tissue lysate (treated with AP)
Lysates/proteins at 10 µg per lane.
Secondary
Lane 1 : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Lane 2 : HRP-conjugate goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 87 kDa
Observed band size: 145 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Pan Trk with purified ab76291 at 1/1000. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Heat mediated antigen retrieval was performed using Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes. Sections were counterstained with hematoxylin. Negative control using PBS instead of primary antibody.
Negative control: No staining on human liver.
The section was incubated with ab76291 for 30 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.
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Unpurified ab76291 staining Pan Trk in murine brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed with formaldehyde, permeabilized with 0.1% Saponin/PBS and blocked with 4% serum for 30 minutes at 25°C, antigen retrieval was by heat mediation with a citrate buffer. Samples were incubated with primary antibody (1/150 in blocking buffer) for 16 hours at 4°C. A FITC-conjugated goat anti-rabbit polyclonal IgG (1/100) was used as the secondary antibody. -
ICC/IF image of Pan Trk staining on culture of mouse DRG neurons using unpurified ab76291 (1/100). The cells were fixed using formaldehyde and permeabilized using 0.2% Triton X-100. The cells were blocked using 10% Goat serum for 1 hour at 22°C. Unpurified ab76291 was diluted 1/100 using PBS and incubated with the cells for 30 mins at 22°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to Alexa Fluor® 488 (1/1000). Neuron was stained using Beta III tubulin antibody (Alexa Fluor® 647)
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 (53)
ab76291 は 53 報の論文で使用されています。
- Yilmaz C et al. ENT-A010, a Novel Steroid Derivative, Displays Neuroprotective Functions and Modulates Microglial Responses. Biomolecules 12:N/A (2022). PubMed: 35327616
- Wuest M et al. Toward in vivo proof of binding of 18F-labeled inhibitor [18F]TRACK to peripheral tropomyosin receptor kinases. EJNMMI Res 12:46 (2022). PubMed: 35907096
- Labib M et al. Tracking the expression of therapeutic protein targets in rare cells by antibody-mediated nanoparticle labelling and magnetic sorting. Nat Biomed Eng 5:41-52 (2021). PubMed: 32719513
- Zeidler M et al. NOCICEPTRA: Gene and microRNA Signatures and Their Trajectories Characterizing Human iPSC-Derived Nociceptor Maturation. Adv Sci (Weinh) 8:e2102354 (2021). PubMed: 34486248
- Chen C et al. Manual Acupuncture at ST37 Modulates TRPV1 in Rats with Acute Visceral Hyperalgesia via Phosphatidylinositol 3-Kinase/Akt Pathway. Evid Based Complement Alternat Med 2021:5561999 (2021). PubMed: 34646326