Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell lysate (ab257594)
製品の概要
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製品名
Human PNP (Nucleoside phosphorylase) knockout HEK-293T cell lysate
Nucleoside phosphorylase キット 製品一覧 -
製品の概要
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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 ab260298 - Human PNP knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
研究分野
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Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
ターゲット情報
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関連疾患
Defects in PNP are the cause of purine nucleoside phosphorylase deficiency (PNP deficiency) [MIM:613179]. It leads to a severe T-cell immunodeficiency with neurologic disorder in children. -
配列類似性
Belongs to the PNP/MTAP phosphorylase family. -
細胞内局在
Cytoplasm > cytoskeleton. - Information by UniProt
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別名
- FLJ94043
- FLJ97288
- FLJ97312
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ab257594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. Predicted molecular weight: 32 kDa.
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特記事項 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa. |
画像
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Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:PNP knockout HeLa cell lysate (20 ug)
Lane 3:Jurkat cell lysate (20 ug)ab109447 was shown to specifically react with Nucleoside phosphorylase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266158 (knockout cell lysate ab257594) was used. Wild-type and Nucleoside phosphorylase knockout samples were subjected to SDS-PAGE. ab109447 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 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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Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:PNP knockout HeLa cell lysate (20 ug)
Lane 3:Jurkat cell lysate (20 ug)
Lane 4:JAR cell lysate (20 ug)ab109559 was shown to specifically react with Nucleoside phosphorylase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266158 (knockout cell lysate ab257594) was used. Wild-type and Nucleoside phosphorylase knockout samples were subjected to SDS-PAGE. ab109559 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 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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Homozygous: 1 bp insertion in exon 2
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)
ab257594 は論文での使用が確認できていません。