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#19526 Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb

 

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CSTコード 包装
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2019年3月26日15時45分 現在
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#19526S100 μL61,000
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#19526T20 μL39,000

Cas9 製品一覧

感度分子量 (kDa)抗体の由来貯法
トランスフェクト150Rabbit 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

すべての種 -
19526 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)免疫組織染色 (パラフィン) (1:400)免疫蛍光細胞染色 (IF-IC) (1:100)免疫蛍光染色 (IF-F) (1:100)フローサイトメトリー (1:800)

下記ステップについては、データシートの右側もあわせてご参照ください。

IHC-P: 抗体希釈液 / 抗原賦活化

CST推奨プロトコールに欠かせない関連製品

特異性・感度
Cas9 (S. pyogenes) (E7M1H) Rabbit mAb recognizes transfected levels of total Cas9 (S. pyogenes) protein. This antibody does not cross-react with Cas9 (S. aureus), FnCpf1, and AsCpf1 proteins.
使用抗原
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu833 of Cas9 (S. Pyogenes) protein.

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社内データ

※下記の社内データは、すべて19526 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. pyogenes), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of Cas9 (S. pyogenes) expression in paraffin-embedded Nprl2-deficient 293 cell pellet (left, positive) or untreated 293 cell pellet (right, negative) using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb. Nprl2 expression was knocked out in the Nprl2-deficient cells by transient transfection of Cas9 (S. pyogenes) and Nprl2-specific guide sequences. 293/Nrpl2 -/- cells were kindly provided by Rachel Wolfson, Lynne Chantranupong, and David Sabatini of MIT, Cambridge, MA.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of Cas9 expression in paraffin-embedded Cas9 (S. Pyogenes) transgenic mouse brain (top-left, positive) and wild type mouse tissues (negative): brain (top-center), colon (top-right), liver (bottom-left), lung (bottom-center), and pancreas (bottom-right), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb.


IF-F

IF-F

Confocal immunofluorescent analysis of brain from constitutive Cas9 (S. pyogenes)-expressing mice (left, positive) or wild type mice (right, negative), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293T cells transiently transfected with a myc-tagged Cas9 (S. pyogenes) construct, using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (green) and Myc-Tag (9B11) Mouse mAb #2276 (red). Colocalization of green and red signals appear yellow in the composite image (bottom-right). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of 293 cells, untransfected (blue) or transfected with Cas9 (S. pyogenes) (green), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (solid lines) or concentraion-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as 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 Streptococcus pyogenes 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).

使用例
 
関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
13079   Immunohistochemistry Application Solutions Kit (Rabbit)
13953   Prestained Protein Marker, Broad Range (11-190 kDa)
14166   Hematoxylin
14177   SignalStain® Mounting Medium
14746   SignalStain® Citrate Unmasking Solution (10X)
15019   Animal-Free Blocking Solution (5X)
2276   Myc-Tag (9B11) Mouse mAb
2278   Myc-Tag (71D10) Rabbit mAb
3900   Rabbit (DA1E) mAb IgG XP® Isotype Control
4412   Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate)
6883   SignalFire™ ECL Reagent
7727   Biotinylated Protein Ladder Detection Pack
8059   SignalStain® DAB Substrate Kit
8112   SignalStain® Antibody Diluent
8114   SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
8961   ProLong® Gold Antifade Reagent with DAPI
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)
9999   Nonfat Dry Milk

Tween is a registered trademark of ICI Americas, Inc.
SignalStain is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
ProLong is a registered trademark of Life Technologies Corporation.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb

Immune Cell Signaling Pathways

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