STAT 5 A/B (pY694/699 + Total) ELISA Kit (ab205715)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 5 µg/ml
- Sample type: Cell culture extracts, Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
製品の概要
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製品名
STAT 5 A/B (pY694/699 + 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% pY694/699 6 2.8% Total 6 3.9% Inter-Assay(日差再現性) サンプル N 平均値 SD CV% pY694/699 3 4.3% Total 3 2.1% -
サンプルの種類
Cell culture extracts, Cell Lysate -
アッセイタイプ
Semi-quantitative -
検出感度
5 µg/ml -
全工程の試験時間
1h 30m -
ステップ
One step assay -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Rat -
製品の概要
Abcam’s STAT5 A/B (pY694/699) and STAT5 A/B (Total) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit (ab204715) is designed for the semi-quantitative measurement of STAT5 A/B (pY694/699) and Total STAT5 A/B protein in Human 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
An alternative fluorescent substrate, ADHP, can be used with this assay. A microplate reader capable of measuring fluorescence is required if using this product.
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
STAT5 A and STAT5 B (collectively known as STAT5 A/B) are two very closely related proteins. Although encoded by different genes, STAT5 A and STAT5 B share 96% identity at the protein level. Similarly to other STAT proteins, STAT5 A and STAT5 B are activated by tyrosine phosphorylation, usually by JAK proteins, at Tyr694 and Tyr699, respectively.
Abcam’s STAT5 A/B (pY694/699) and STAT5 A/B (Total) SimpleStep ELISA® kit (ab204715) has been re-developed with new detection antibody. From February 18, 2016 the kit will be delivered with a new detection total STAT5 antibody. This change was necessary because of a decrease in the performance of the polyclonal STAT5 antibody that was previously used for capture in recent production lots. We have identified a new antibody to use in the SimpleStep ELISA platform that provides higher performance.
Abcam 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
Pre-coated microplate (12 x 8 well strips)
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 10X Wash Buffer PT 1 x 15ml 50X Cell Extraction Enhancer Solution 1 x 1ml 5X Cell Extraction Buffer PTR 1 x 10ml Lyophilized STAT5 A/B Control Lysate 1 vial Plate Seal 1 unit SimpleStep Pre-Coated 96 Well Microplate (12 x 8 well strips) 1 unit STAT5 A/B (pY694/699) Capture Antibody 1 x 1.5ml STAT5 A/B (pY694/699) Detector Antibody 1 x 1.5ml STAT5 A/B (Total) Capture Antibody 1 x 1.5ml STAT5 A/B (Total) Detector Antibody 1 x 1.5ml Stop Solution 1 x 12ml TMB Substrate 1 x 12ml -
研究分野
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細胞内局在
STAT5a + STAT5b: Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. STAT5 (Total): Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. -
参照データベース
- Entrez Gene: 6776 Human
- Entrez Gene: 6777 Human
- Entrez Gene: 20850 Mouse
- Entrez Gene: 20850 Mouse
- Entrez Gene: 20851 Mouse
- Entrez Gene: 24918 Rat
- Entrez Gene: 24918 Rat
- Entrez Gene: 25126 Rat
see all
関連製品
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画像
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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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Cell line analysis for Total STAT5 A/B 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 STAT5 A/B (pY694/699) phosphorylation in A431 cells in response to EGF treatment. A431 cells were cultured in 96-well tissue culture plates and treated (15 min) with a dose-range of EGF before cell lysis. Data from quadruplicate measurements of STAT5 A/B (pY694/699) are plotted and compared against Total STAT5 A/B levels.
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Example of a typical STAT5 A/B (pY694/699) and STAT5 A/B (Total) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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 (2)
ab205715 は 2 報の論文で使用されています。
- Moore J et al. Random errors in protein synthesis activate an age-dependent program of muscle atrophy in mice. Commun Biol 4:703 (2021). PubMed: 34103648
- Tong JJ et al. Effect of 17ß-estradiol on milk production, hormone secretion, and mammary gland gene expression in dairy cows. J Dairy Sci 101:2588-2601 (2018). PubMed: 29274969