Human DCP1A knockout HEK-293 cell lysate (ab261665)
製品の概要
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製品名
Human DCP1A knockout HEK-293 cell lysate -
製品の概要
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
HEK-293 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 7 bp deletion; Frameshift = 100% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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特記事項
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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アプリケーション
適用あり: WBmore details
製品の特性
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保存方法
Store at -80°C. Please refer to protocols. -
内容 1 kit ab280415 - Human DCP1A knockout HEK293 cell lysate 1 x 100µg ab259780 - Human wild-type HEK-293 cell lysate 1 x 100µg -
研究分野
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Cell type
epithelial -
Gender
Female
ターゲット情報
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機能
Necessary for the degradation of mRNAs, both in normal mRNA turnover and in nonsense-mediated mRNA decay. Removes the 7-methyl guanine cap structure from mRNA molecules, yielding a 5'-phosphorylated mRNA fragment and 7m-GDP. Contributes to the transactivation of target genes after stimulation by TGFB1. -
組織特異性
Detected in heart, brain, placenta, lung, skeletal muscle, liver, kidney and pancreas. -
配列類似性
Belongs to the DCP1 family. -
細胞内局在
Cytoplasm > P-body. Nucleus. Co-localizes with NANOS3 in the processing bodies (By similarity). Predominantly cytoplasmic, in processing bodies (PB). Nuclear, after TGFB1 treatment. Translocation to the nucleus depends on interaction with SMAD4. - Information by UniProt
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別名
- DCP1 decapping enzyme homolog A
- Dcp1a
- DCP1A_HUMAN
see all
関連製品
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KO cell lines
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Related Products
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ab261665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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WB |
Use at an assay dependent concentration.
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特記事項 |
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WB
Use at an assay dependent concentration. |
画像
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Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2: DCP1A knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lanes 1 - 3: Merged signal (red and green). Green - ab183709 observed at 75 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab183709 was shown to specifically react with DCP1A in wild-type HEK-293 cells as signal was lost in DCP1A knockout cell line ab261857 (knockout cell lysate ab261665). Wild-type and DCP1A knockout samples were subjected to SDS-PAGE. Ab183709 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 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. -
Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2: DCP1A knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lanes 1 - 3: Merged signal (red and green). Green - ab47811 observed at 75 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab47811 was shown to recognize DCP1A in wild-type HEK-293 cells as signal was lost at the expected MW in DCP1A knockout cell line ab261857 (knockout cell lysate ab261665). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and DCP1A knockout samples were subjected to SDS-PAGE. Ab47811 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml 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. -
X = 7 bp deletion
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
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 (0)
ab261665 は論文での使用が確認できていません。