Revision 2
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P, IF-IC, FC-FP

REACTIVITY:

All

SENSITIVITY:

Transfected Only

MW (kDa):

150

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q99ZW2

Entrez-Gene Id:

901176

Product Information

Product Usage Information

This product is the carrier free version of product #19526. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #19526

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

Cas9 (S. pyogenes) (E7M1H) Rabbit mAb (BSA and Azide Free) recognizes transfected levels of total Cas9 (S. pyogenes) protein. This antibody does not cross-react with Cas9 (S. aureus), FnCpf1, and AsCpf1 proteins.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu833 of Cas9 (S. Pyogenes) protein.

Background

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 extrachromosomal 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).

  1. Horvath, P. and Barrangou, R. (2010) Science 327, 167-70.
  2. Wiedenheft, B. et al. (2012) Nature 482, 331-8.
  3. Singh, P. et al. (2015) Genetics 199, 1-15.
  4. Cong, L. et al. (2013) Science 339, 819-23.
  5. Mali, P. et al. (2013) Science 339, 823-6.
  6. Li, D. et al. (2013) Nat Biotechnol 31, 681-3.
  7. Shen, B. et al. (2013) Cell Res 23, 720-3.
  8. Niu, Y. et al. (2014) Cell 156, 836-43.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 2
#98605

Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (BSA and Azide Free)

Western Blotting Image 1: Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Cas9 (S. pyogenes) (E7M1H) XP® 兔单克隆抗体(上图)和 Myc-Tag (71D10) 兔单克隆抗体 #2278(下图)对转染空载 (-) 或转染表达 Cas9(酿脓链球菌)的表达载体的 293T 细胞的提取物进行蛋白印迹分析。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 1: Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Cas9 (S. pyogenes) (E7M1H) XP® 兔单克隆抗体 对在石蜡包埋的 Cas9(酿脓链球菌)转基因小鼠脑细胞(左上图,阳性)和脑(顶部中间)、结肠(右上图)、肝(左下图)、肺(底部中间)和胰腺(右下图)等野生型小鼠组织(阴性)中表达的 Cas9 进行免疫组织化学分析。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 2: Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Cas9 (S. pyogenes) (E7M1H) XP® 兔单克隆抗体 对在石蜡包埋的、Nprl2 缺陷型 293 细胞沉淀物(左图,阳性)或未处理的 293 细胞沉淀物(右图,阴性)中表达的 Cas9(酿脓链球菌)进行免疫组织化学分析。通过瞬时转染 Cas9(酿脓链球菌)和 Nprl2 特异性引导序列,敲除 Nprl2 缺陷型细胞中的 Nprl2 表达。293/Nrpl2 -/- 细胞由马萨诸塞州坎布里奇麻省理工学院的 Rachel Wolfson 和 Lynne Chantranupong 惠赠。数据的生成使用的是该产品的常规组分产品。
Immunofluorescence Image 1: Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Cas9 (S. pyogenes) (E7M1H) XP® 兔单克隆抗体(绿色)和 Myc-Tag (9B11) Mouse 单克隆抗体 #2276(红色)对瞬时转染带有 myc 标签的 Cas9(酿脓链球菌)表达载体的 293T 细胞进行共聚焦免疫荧光分析。绿色和红色信号共定位在合成图像(右下图)中显示为黄色。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。数据的生成使用的是该产品的常规组分产品。
Flow Cytometry Image 1: Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Cas9 (S. pyogenes) (E7M1H) XP® 兔单克隆抗体(实线)或浓度匹配的兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900(虚线)对未转染(蓝色)或转染 Cas9(酿脓链球菌(绿色)的 293 细胞进行流式细胞分析。抗兔 IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 偶联物) #4412 用作二抗。数据的生成使用的是该产品的常规组分产品。