Anti-ERK1 抗体 [EP4967] (ab109282)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP4967] to ERK1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-ERK1 antibody [EP4967]
ERK1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP4967] to ERK1 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore 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: HEK-293T, Jurkat, A375, A431 and HUVEC cell lysates. IHC-P: Human colon adenocarcinoma tissue. Flow Cyt (intra): Jurkat cells, Hap1 cells.
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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: 40% Glycerol (glycerin, glycerine), 59% PBS, 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
EP4967 -
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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Isotype control
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KO cell lines
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KO cell lysates
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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ab109282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) |
1/30.
ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 44 kDa.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. Heat up to 98°C, below boiling, and then let cool for 10-20 min. |
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ICC/IF |
1/100.
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特記事項 |
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Flow Cyt (Intra)
1/30. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Predicted molecular weight: 44 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Heat up to 98°C, below boiling, and then let cool for 10-20 min. |
ICC/IF
1/100. |
ターゲット情報
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機能
Involved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK-1. Phosphorylates EIF4EBP1; required for initiation of translation. Phosphorylates microtubule-associated protein 2 (MAP2). Phosphorylates SPZ1 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4). -
配列類似性
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain. -
ドメイン
The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases. -
翻訳後修飾
Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Dephosphorylated by PTPRJ at Tyr-204. - Information by UniProt
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参照データベース
- Entrez Gene: 5595 Human
- Entrez Gene: 26417 Mouse
- Entrez Gene: 50689 Rat
- Omim: 601795 Human
- SwissProt: P27361 Human
- SwissProt: Q63844 Mouse
- SwissProt: P21708 Rat
- Unigene: 861 Human
see all -
別名
- ERK 1 antibody
- ERK-1 antibody
- ERK1 antibody
see all
画像
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Flow cytometry overlay histogram showing wild-type Hap1 (green line) and MAPK3 knockout Hap1 stained with ab109282 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab109282) (1x 106 in 100μl at 0.2 μg/ml (1/11400)) 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 Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type Hap1 - black line, MAPK3 knockout Hap1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
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-ERK1 antibody [EP4967] (ab109282) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : MAPK3 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab109282 observed at 43 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109282 was shown to react with ERK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266519 (knockout cell lysate ab257099) was used. Wild-type HEK-293T and MAPK3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109282 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling ERK1 with purified ab109282 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/Immunofluorescence analysis of Jurkat (human acute T cell leukemia) cells labelling ERK1 with purified ab109282 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
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Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling ERK1 with purified ab109282 at 1/30 dilution (red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
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Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Green signal from target - ab109282 observed at 42 kDa
Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
ab109282 was shown to specifically react with ERK1 when ERK1 knockout samples were used.
Wild-type and ERK1 knockout samples were subjected to SDS-PAGE. ab109282 and ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Predicted band size : 43 kDa
Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Green signal from target
Lanes 3 and 4: Red signal from loading control
Lanes 5 and 6: Merged (red and green) signalRed - loading control, ab8226 observed at 42 kDa or ab8245, observed at 37 kDa
This western blot image is a comparison between ab109282 and a competitor's top cited rabbit polyclonal antibody.
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All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/10000 dilution (purified)
Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 44 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/10000 dilution (purified)
Lane 1 : Jurkat cell lysate
Lane 2 : A375 cell lysate
Lane 3 : HUVEC cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 44 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/1000 dilution (unpurified)
Lane 1 : Jurkat cell lysate
Lane 2 : A375 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HUVEC cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 44 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colonic adenocarcinoma tissue labelling ERK1 with unpurified ab109282 at 1/100 dilution.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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 (7)
ab109282 は 7 報の論文で使用されています。
- Yu Y et al. The lncRNA KIF9-AS1 Accelerates Hepatocellular Carcinoma Growth by Recruiting DNMT1 to Promote RAI2 DNA Methylation. J Oncol 2022:3888798 (2022). PubMed: 36276278
- Gao Z et al. Analysis of RIOK2 Functions in Mediating the Toxic Effects of Deoxynivalenol in Porcine Intestinal Epithelial Cells. Int J Mol Sci 23:N/A (2022). PubMed: 36361502
- Fan H et al. Chromatin Accessibility and Transcriptomic Alterations in Murine Ovarian Granulosa Cells upon Deoxynivalenol Exposure. Cells 10:N/A (2021). PubMed: 34831041
- Wang Y et al. TGF-β1/SMOC2/AKT and ERK axis regulates proliferation, migration, and fibroblast to myofibroblast transformation in lung fibroblast, contributing with the asthma progression. Hereditas 158:47 (2021). PubMed: 34876240
- Liu D et al. Long non-coding RNA GASL1 restrains gastric carcinoma cell proliferation and metastasis by sponging microRNA-106a. Cell Cycle 19:2611-2621 (2020). PubMed: 32897806
- Zhi Q et al. Podocalyxin-like protein promotes gastric cancer progression through interacting with RUN and FYVE domain containing 1 protein. Cancer Sci 110:118-134 (2019). PubMed: 30407695
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371