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#51610 Cas9 (S. aureus) (E4G3U) Rabbit mAb

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2019年1月15日15時15分 現在
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#51610S100 μL56,000
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Cas9 製品一覧 | #51610 が入っているAntibody Sampler キット一覧

感度分子量 (kDa)抗体の由来貯法
トランスフェクト124Rabbit IgG-20℃
種交差性 (社内試験済)
交差する可能性がある種 i

社内試験はしていませんが、配列が100%相同であるため反応すると推定される種

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51610 の推奨プロトコール i

最適な結果を得るために:Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。
推奨プロトコールはCST社内試験の徹底的なバリデーションに基づいて作成されておりますので、正確かつ再現性の高い結果が得られます。

注:各製品に最適化されたプロトコールをリンクしています。

 

ウェスタンブロッティング (1:1000)免疫蛍光細胞染色 (IF-IC) (1:200)フローサイトメトリー (1:100)

CST推奨プロトコールに欠かせない関連製品

特異性・感度
トランスフェクトレベルのCas9 (S. aureus) タンパク質を検出します。Cas9 (S. pyogenes)、AsCpf1 (Strain BV3L6)、FnCpf (Strain U112) タンパク質とは交差しません。
使用抗原
Cas9 (S. aureus) タンパク質のVal905 周辺領域 (合成ペプチド)

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社内データ

※下記の社内データは、すべて51610 の推奨プロトコールで実験した結果です。

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Cas9 (S. aureus), Cas9 (S. pyogenes), AsCpf1 (Strain BV3L6), or FnCpf1 (Strain U112) (+), using Cas9 (S. aureus) (E4G3U) Rabbit mAb (upper) and Myc-Tag (7110) Rabbit mAb #2278 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293T cells transfected with a construct expressing myc-tagged Cas9 (S. aureus) using Cas9 (S. aureus) (E4G3U) Rabbit mAb (green) and Myc-Tag (9B11) Mouse mAb #2276 (red). Overlapping signals are shown in yellow. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of 293T cells, untransfected (blue) or transfected with a construct expressing myc-tagged Cas9 (S. aureus) (green), using Cas9 (S. aureus) Rabbit mAb (solid line) compared to a concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.


バックグラウンド

The CRISPR associated protein 9 (Cas9) is an RNA-guided DNA nuclease and part of the CRISPR antiviral immunity system that provides adaptive immunity against extra chromosomal genetic material (1). The CRISPR antiviral mechanism of action involves three steps: (i), acquisition of foreign DNA by host bacterium; (ii), synthesis and maturation of CRISPR RNA (crRNA), followed by the formation of RNA-Cas nuclease protein complexes; and (iii), target interference through recognition of foreign DNA by the complex and its cleavage by Cas nuclease activity (2). The type II CRISPR/Cas antiviral immunity system provides a powerful tool for precise genome editing and has potential for specific gene regulation and therapeutic applications (3). The Cas9 protein and a guide RNA consisting of a fusion between a crRNA and a trans-activating crRNA (tracrRNA) must be introduced or expressed in a cell. A 20-nucleotide sequence at the 5' end of the guide RNA directs Cas9 to a specific DNA target site. As a result, Cas9 can be "programmed" to cut various DNA sites both in vitro and in cells and organisms. CRISPR/Cas9 genome editing tools have been used in many organisms, including mouse and human cells (4,5). Research studies demonstrate that CRISPR can be used to generate mutant alleles or reporter genes in rodents and primate embryonic stem cells (6-8).

Cas9 (S. aureus) is a Cas9 ortholog that is smaller, but as efficient, as the more commonly used Cas9 ortholog, Cas9 (S. Pyogenes) (9).

使用例
 
関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
13953   Prestained Protein Marker, Broad Range (11-190 kDa)
2276   Myc-Tag (9B11) Mouse mAb
2729   Normal Rabbit IgG
3900   Rabbit (DA1E) mAb IgG XP® Isotype Control
4084   DRAQ5®
4412   Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate)
5127   Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate)
6883   SignalFire™ ECL Reagent
7727   Biotinylated Protein Ladder Detection Pack
9863   Protein A Agarose Beads
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)
9999   Nonfat Dry Milk

The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation, for research use only for immunocytometry, immunohistochemistry, high content screening (HCS) analysis, or flow cytometry applications.
DRAQ5 is a registered trademark of Biostatus Limited.
Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
The transfer of this product is contingent on the buyer using the purchased product solely in research conducted by the buyer (whether the buyer is an academic or for-profit entity), for Immunocytochemistry, high content screening (HCS) analysis, or flow cytometry applications. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) resale, whether or not such product or its components are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

本製品は試験研究用です。

Cas9 (S. aureus) (E4G3U) Rabbit mAb

Metabolic Reprogramming in Disease

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