ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) + Total ELISA Kit (ab176660)
Key features and details
- One-wash 90 minute protocol
- Sample type: Cell Lysate, Tissue Homogenate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Mouse, Human
製品の概要
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製品名
ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) + Total ELISA Kit -
検出方法
Colorimetric -
再現性
Intra-Assay(同時再現性) サンプル N 平均値 SD CV% (pT202/Y204) 6 3.3% (Total) 6 3% Inter-Assay(日差再現性) サンプル N 平均値 SD CV% (pT202/Y204) 3 1.3% (Total) 3 4.9% -
サンプルの種類
Cell Lysate, Tissue Homogenate -
アッセイタイプ
Semi-quantitative -
全工程の試験時間
1h 30m -
ステップ
One step assay -
種交差性
交差種: Mouse, Human
交差が予測される動物種: Rat -
製品の概要
Abcam’s ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) + Total ELISA kit is designed for the semi-quantitative measurement of ERK1/2 (pT202/Y204) and Total ERK1/2 protein in Human and mouse cells.
The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
ASSAY SPECIFICITY
The ERK1/2 (pT202/Y204) assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737.2 [ERK1], NP 620407 [ERK2]) in cellular lysates, only when phosphorylated at Thr202/Tyr204.
The ERK1/2 Total assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737.2 [ERK1], NP 620407 [ERK2]) in cellular lysates, irrespective of phosphorylation status.
SPECIES REACTIVITY
This kit detects ERK1/2 (pT202/Y204) and ERK1/2 Total in Human and mouse cell culture extracts. Detection in rat samples is also expected. Other species should be tested on a case-by-case basis.
Serum and plasma samples have not been tested with this kit.
As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.
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特記事項
Estimated sensitivity: Phospho-ERK (Thr202/Tyr204): 0.1 ng/mL (tested with recombinant protein), Total ERK1/2: 0.6 ng/mL (tested with recombinant protein)
Range: Phospho-ERK (Thr202/Tyr204): 0.2-20ng/mL, Total ERK1/2: 0.6-60 ng/mLAbcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
試験プラットフォーム
Microplate
製品の特性
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保存方法
Store at +4°C. Please refer to protocols. -
内容 1 x 96 tests 1 x 96 tests 10X Wash Buffer PT 1 x 15ml 1 x 15ml 50X Cell Extraction Enhancer Solution 1 x 1ml 1 x 1ml 5X Cell Extraction Buffer PTR 1 x 10ml 1 x 10ml ERK1/2 (pT202/Y204) Capture Antibody 1 x 1.5ml 1 x 1.5ml ERK1/2 (pT202/Y204) Detector Antibody 1 x 1.5ml 1 x 1.5ml ERK1/2 (Total) Capture Antibody 1 x 1.5ml 1 x 1.5ml ERK1/2 (Total) Detector Antibody 1 x 1.5ml 1 x 1.5ml Lyophilized ERK1/2 Control Lysate 1 vial 1 vial Plate Seal 1 unit 1 unit SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit 1 unit Stop Solution 1 x 12ml 1 x 12ml TMB Substrate 1 x 12ml 1 x 12ml -
研究分野
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細胞内局在
ERK2: Nucleus. -
別名
- ERK
- ERK 1
- ERK 2
see all -
参照データベース
- Entrez Gene: 5594 Human
- Entrez Gene: 5595 Human
- Entrez Gene: 26413 Mouse
- Entrez Gene: 26417 Mouse
- Entrez Gene: 116590 Rat
- Entrez Gene: 50689 Rat
- Omim: 176948 Human
- Omim: 601795 Human
see all
画像
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The ERK1/2 lysate dilution series was prepared as described. Raw data values are shown in the table.
Kit Control lysates are provided at a concentration that give consistent signal between different lots. Lysates are produced and formulated by signal intensity to be consistent to within 30% of the previous lot. As such, Control lysates are not provided with a protein concentration.
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The ERK1/2 lysate dilution series was prepared as described. Raw data values are shown in the table.
Kit Control lysates are provided at a concentration that give consistent signal between different lots. Lysates are produced and formulated by signal intensity to be consistent to within 30% of the previous lot. As such, Control lysates are not provided with a protein concentration.
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Example of a typical ERK1/2 (pT202/Y204) and ERK1/2 (Total) recombinant protein standard curve. The proportion of total protein that is phosphorylated is unknown - data is indicative only. Background-subtracted data values (mean +/- SD) are graphed.
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Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of ERK1/2 (pT202/Y204) are normalized and plotted.
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Cell line analysis for Total ERK1/2 from 100 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
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Induction of ERK1/2 (pT202/Y204) phosphorylation in MCF-7 cells in response to EGF treatment. MCF-7 cells were cultured in 96-well tissue culture plates, serum starved and treated (10 min) with a dose-range of EGF before cell lysis. Data from quadruplicate measurements of ERK1/2 (pT202/Y204) are plotted and compared against Total ERK1/2 protein levels. Comparative ERK1/2 (pT202/Y204) and ERK1/2 (Total) data also shown by Western Blot.
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SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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To learn more about the advantages of SimpleStep ELISA® kits see here.
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 (33)
ab176660 は 33 報の論文で使用されています。
- Wang M et al. BMP4 preserves the developmental potential of mESCs through Ube2s- and Chmp4b-mediated chromosomal stability safeguarding. Protein Cell 13:580-601 (2022). PubMed: 35147915
- Younis NS & Ghanim AMH The Protective Role of Celastrol in Renal Ischemia-Reperfusion Injury by Activating Nrf2/HO-1, PI3K/AKT Signaling Pathways, Modulating NF-κb Signaling Pathways, and Inhibiting ERK Phosphorylation. Cell Biochem Biophys 80:191-202 (2022). PubMed: 35157199
- Fawzy MA et al. Vincamine Modulates the Effect of Pantoprazole in Renal Ischemia/Reperfusion Injury by Attenuating MAPK and Apoptosis Signaling Pathways. Molecules 27:N/A (2022). PubMed: 35209172
- Li Y & Dou S FLOT2 Promotes the Proliferation and Epithelial-mesenchymal Transition of Cervical Cancer by Activating the MEK/ERK1/2 Pathway. Balkan Med J 39:267-274 (2022). PubMed: 35872627
- Xie MZ et al. Proteomics-based evaluation of the mechanism underlying vascular injury via DNA interstrand crosslinks, glutathione perturbation, mitogen-activated protein kinase, and Wnt and ErbB signaling pathways induced by crotonaldehyde. Clin Proteomics 19:33 (2022). PubMed: 36002804