Revision 5

#13202Store 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, IP, IF-IC, FC-FP

REACTIVITY:

All

SENSITIVITY:

Transfected Only

MW (kDa):

Source/Isotype:

Rabbit IgG

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:500 - 1:2000
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

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 #58009.

Specificity / Sensitivity

V5-Tag (D3H8Q) Rabbit mAb recognizes transfected levels of recombinant protein containing the V5 epitope tag.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic V5-tag peptide.

Background

Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein’s biochemical properties.

The V5 tag is a 14 amino acid peptide derived from a small epitope (Pk) on the P and V proteins of simian virus 5 (SV5), a member of the paramyxovirus family (1). The V5 tag can be used to detect expression of recombinant proteins in bacteria, yeast, insects, and mammalian cell systems.

  1. Hanke, T. et al. (1995) J Virol Methods 53, 149-56.

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™ IP: Immunoprecipitation 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 5
#13202

V5-Tag (D3H8Q) Rabbit mAb

Western Blotting Image 1: V5-Tag (D3H8Q) Rabbit mAb Expand Image
使用 V5-Tag (D3H8Q) Rabbit mAb 和 β-Actin (D6A8) Rabbit mAb #8457 对未转染 (-) 或转染带有 V5 标记的融合蛋白 (+) 的 COS-7 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 1: V5-Tag (D3H8Q) Rabbit mAb Expand Image

使用 V5-Tag (D3H8Q) Rabbit mAb #13202 对来自 COS-7 +V5-N-Ras 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。

Immunoprecipitation Image 1: V5-Tag (D3H8Q) Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(泳道 2)或 V5-Tag (D3H8Q) Rabbit mAb(泳道 3)对 COS-7 细胞提取物经转染带有 V5 标记的融合蛋白进行免疫沉淀。泳道 1 是 10% 输入对照。使用 V5-Tag (D3H8Q) Rabbit mAb 进行蛋白质印迹分析。
Immunofluorescence Image 1: V5-Tag (D3H8Q) Rabbit mAb Expand Image
使用 V5-Tag (D3H8Q) Rabbit mAb(绿色)、DyLight 554 Phalloidin #13054(红色)和 DRAQ5® #4084(伪彩蓝色),对经 V5 标记的构建体转染(左图)或空载转染(右图)的 COS-7 细胞进行共聚焦免疫荧光分析。
Flow Cytometry Image 1: V5-Tag (D3H8Q) Rabbit mAb Expand Image
使用 V5-Tag (D3H8Q) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对转染空载(蓝色)或转染带有 V5 标记的结构(绿色)的 293T 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。