Revision 12

#2276Store 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, IHC-P, IF-IC, FC-FP, ChIP

REACTIVITY:

All

SENSITIVITY:

Transfected Only

MW (kDa):

Source/Isotype:

Mouse IgG2a kappa

Product Information

Product Usage Information

For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:250
Immunohistochemistry (Paraffin) 1:1500 - 1:6000
Immunofluorescence (Immunocytochemistry) 1:1000 - 1:2000
Flow Cytometry (Fixed/Permeabilized) 1:500 - 1:2000
Chromatin IP 1:50

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 #58730.

Specificity / Sensitivity

Myc-Tag (9B11) Mouse mAb detects exogenously expressed proteins containing the Myc epitope tag. This antibody recognizes the Myc tag fused to either the amino or carboxy terminus of targeted proteins in transfected cells. Myc-Tag (9B11) Mouse mAb detects exogenously expressed Myc-tagged proteins in cells expressed under a CMV promoter. Expression under other promoters has not been evaluated.
The antibody may cross-react with c-myc protein. The antibody may weakly cross-react with a protein of unknown origin ~90kDa.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues 410-419 of human c-Myc (EQKLISEEDL).

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 Myc epitope tag is widely used to detect expression of recombinant proteins in bacteria, yeast, insect and mammalian cell systems (1).

  1. Munro, S. and Pelham, H.R. (1984) EMBO J. 3, 3087-3093.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP

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.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered 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 12
#2276

Myc-Tag (9B11) Mouse mAb

Western Blotting Image 1: Myc-Tag (9B11) Mouse mAb Expand Image
使用 Myc-Tag (9B11) Mouse mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染空载(泳道 1)或瞬时转染编码带有 Myc/DDK 标签的 SARS-CoV-2 RNA 依赖性 RNA 聚合酶蛋白(泳道 2)的 293T 细胞提取物进行蛋白质印迹法分析。
Western Blotting Image 2: Myc-Tag (9B11) Mouse mAb Expand Image
使用 Myc-Tag (9B11) Mouse mAb 对不同浓度的重组人 SYN3-MYC/DYKDDDDK 进行蛋白质印迹法分析。
Western Blotting Image 1: Myc-Tag (9B11) Mouse mAb Expand Image
Simple Western™ analysis of lysates (0.1 mg/mL) from 293T cells, transfected with a construct expressing Myc/DDK tagged MMP-8 using Myc-Tag (9B11) Mouse mAb #2276. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的单一靶标条带(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: Myc-Tag (9B11) Mouse mAb Expand Image
对 293T 转染细胞提取物的 SARS-CoV-2 Spike-Myc/DDK 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Mouse (E5Y6Q) mAb IgG2a Isotype Control #61656,泳道 3 为 Myc-Tag (9B11) Mouse mAb。使用 Myc-Tag (71D10) Rabbit mAb #2278 进行蛋白质印迹分析。
Immunohistochemistry Image 1: Myc-Tag (9B11) Mouse mAb Expand Image
使用 Myc-Tag (9B11) Mouse mAb,对对照(左)或羧基末端 Myc 标记的蛋白(右)转染的石蜡包埋的 COS 细胞沉淀物进行免疫组织化学分析。
Immunofluorescence Image 1: Myc-Tag (9B11) Mouse mAb Expand Image
使用 Myc-Tag (9B11) Mouse mAb(绿色)对转染带 Myc 标记蛋白的 COS 细胞进行共聚焦免疫荧光分析。肌动蛋白丝用 DyLight 554 Phalloidin #13054(红色)标记。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: Myc-Tag (9B11) Mouse mAb Expand Image
使用 Myc-Tag (9B11) Mouse mAb(实线)或浓度匹配的 Mouse (G3A1) mAb IgG1 Isotype Control #5415(虚线)对转染空载(蓝色)或转染带有 Myc 标记的表达载体(绿色)的 293T 细胞进行流式细胞分析。Anti-mouse IgG (H+L),F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 作为二抗。
Chromatin Immunoprecipitation Image 1: Myc-Tag (9B11) Mouse mAb Expand Image
293T 细胞未转染(左小图)或转染 Myc 标签的人 Stat3 表达载体(右小图),然后用 Human Interleukin-6 (hIL-6) #8904(100 ng/ml,30 分钟)进行处理。使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对细胞中提取的交联染色质和 Myc-Tag (9B11) Mouse mAb #2276、Stat3α (D1A5) XP® Rabbit mAb #8768 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀法分析。使用 SimpleChIP® Human c-Fos Promoter Primers #4663、human IRF-1 promoter primers 和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 对富集的 DNA 进行定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。