Revision 7

#4970Store at -20C

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, W-S, IHC-P, IF-F, IF-IC, FC-FP

REACTIVITY:

H M R Mk B Pg

SENSITIVITY:

Endogenous

MW (kDa):

45

Source/Isotype:

Rabbit IgG

UniProt ID:

#P60709

Entrez-Gene Id:

60

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Frozen) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #93473.

Specificity / Sensitivity

β-Actin (13E5) Rabbit mAb detects endogenous levels of total β-actin protein. Depsite the high sequence identity between the cytoplasmic actin isoforms, β-actin and cytoplasmic γ-actin, β-Actin (13E5) Rabbit mAb #4970 does not cross-react with cytoplasmic γ-actin, or any other actin isoforms.

Species Reactivity:

Human, Mouse, Rat, Monkey, Bovine, Pig

Species predicted to react based on 100% sequence homology

Hamster, Chicken, Dog, Horse, Rabbit

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino-terminus of human β-actin protein.

Background

Actin, a ubiquitous eukaryotic protein, is the major component of the cytoskeleton. At least six isoforms are known in mammals. Nonmuscle β- and γ-actin, also known as cytoplasmic actin, are ubiquitously expressed, controlling cell structure and motility (1). While all actin isoforms are highly homologous, cytoplasmic β- and γ-actin protein sequences differ by only four biochemically similar amino acids (2). For this reason, antibodies raised to β-actin may cross-react with γ-actin, and vice versa. α-cardiac and α-skeletal actin are expressed in striated cardiac and skeletal muscles, respectively; two smooth muscle actins, α- and γ-actin, are found primarily in vascular smooth muscle and enteric smooth muscle, respectively. These actin isoforms regulate the contractile potential of muscle cells (1). Actin exists mainly as a fibrous polymer, F-actin. In response to cytoskeletal reorganizing signals during processes such as cytokinesis, endocytosis, or stress, cofilin promotes fragmentation and depolymerization of F-actin, resulting in an increase in the monomeric globular form, G-actin (3). The ARP2/3 complex stabilizes F-actin fragments and promotes formation of new actin filaments (3). Research studies have shown that actin is hyperphosphorylated in primary breast tumors (4). Cleavage of actin under apoptotic conditions has been observed in vitro and in cardiac and skeletal muscle, as shown in research studies (5-7). Actin cleavage by caspase-3 may accelerate ubiquitin/proteasome-dependent muscle proteolysis (7).

  1. Herman, I.M. (1993) Curr. Opin. Cell Biol. 5, 48-55.
  2. Perrin, B.J. and Ervasti, J.M. (2010) Cytoskeleton (Hoboken) 67, 630-4.
  3. Condeelis, J. (2001) Trends Cell Biol 11, 288-93.
  4. Lim, Y.P. et al. (2004) Clin Cancer Res 10, 3980-7.
  5. Kayalar, C. et al. (1996) Proc Natl Acad Sci U S A 93, 2234-8.
  6. Communal, C. et al. (2002) Proc Natl Acad Sci U S A 99, 6252-6.
  7. Du, J. et al. (2004) J Clin Invest 113, 115-23.

Species Reactivity

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

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) 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.
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 7
#4970

β-Actin (13E5) Rabbit mAb

Western Blotting Image 1: β-Actin (13E5) Rabbit mAb Expand Image
使用 beta-Actin (13E5) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
Western Blotting Image 2: β-Actin (13E5) Rabbit mAb Expand Image
使用 β-Actin (13E5) Rabbit Mab(上图)和 Pan-Actin Antibody # 对重组肌动蛋白同工型进行蛋白质印迹分析4968 (下图)。
Western Blotting Image 1: β-Actin (13E5) Rabbit mAb Expand Image
使用 ß-Actin (13E5) Rabbit mAb #4970 对 HeLa 细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunohistochemistry Image 1: β-Actin (13E5) Rabbit mAb Expand Image
使用 beta-Actin (13E5) Rabbit mAb 对石蜡包埋的人心脏细胞进行免疫组织化学分析。注意心脏肌动蛋白缺乏染色。
Immunohistochemistry Image 2: β-Actin (13E5) Rabbit mAb Expand Image
使用 beta-Actin (13E5) Rabbit mAb 对石蜡包埋的人平滑肌瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: β-Actin (13E5) Rabbit mAb Expand Image
使用 β-actin (13E5) Rabbit mAb 对石蜡包埋的 4T1 同基因小鼠肿瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: β-Actin (13E5) Rabbit mAb Expand Image
在对照肽(左)或 beta-Actin Blocking Peptide #1025(右)存在的情况下,使用 beta-Actin (13E5) Rabbit mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 5: β-Actin (13E5) Rabbit mAb Expand Image
使用 beta-Actin (13E5) Rabbit mAb 对石蜡包埋的人骨骼肌细胞进行免疫组织化学分析。注意骨骼肌肌动蛋白缺乏染色。
Immunofluorescence Image 1: β-Actin (13E5) Rabbit mAb Expand Image
使用 β-Actin (13E5) Rabbit mAb #4970(红色)和 DAPI #4083(蓝色)在低放大倍率(左图)和高放大倍率(右图)下对固定冷冻的小鼠胰腺进行共聚焦免疫荧光分析。
Immunofluorescence Image 1: β-Actin (13E5) Rabbit mAb Expand Image
使用 β-Actin (13E5) Rabbit mAb(绿色)对 COS-7 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5®#4084(DNA 荧光染料)。
Flow Cytometry Image 1: β-Actin (13E5) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 β-Actin (13E5) Rabbit mAb(实线)对 HeLa 细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。