Revision 3

#76266Store 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, IP, ChIP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

62

Source/Isotype:

Rabbit IgG

UniProt ID:

#P12524

Entrez-Gene Id:

4610

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
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 -20ºC. Do not aliquot the antibody.

Specificity / Sensitivity

L-Myc (E3M5P) Rabbit mAb recognizes endogenous levels of total L-Myc protein. Cross-reactivity was not observed with other family members.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human L-Myc protein. The epitope has been mapped to residues surrounding Thr190.

Background

Members of the Myc/Max/Mad network function as transcriptional regulators with roles in various aspects of cell behavior, including proliferation, differentiation, and apoptosis (1). These proteins share a common basic-helix-loop-helix leucine zipper (bHLH-ZIP) motif required for dimerization and DNA-binding. Max was originally discovered based on its ability to associate with c-Myc and found to be required for the ability of Myc to bind DNA and activate transcription (2). Subsequently, Max has been viewed as a central component of the transcriptional network, forming homodimers as well as heterodimers with other members of the Myc and Mad families (1). The association between Max and either Myc or Mad can have opposing effects on transcriptional regulation and cell behavior (1). The Mad family consists of four related proteins; Mad1, Mad2 (Mxi1), Mad3, and Mad4, and the more distantly related members of the bHLH-ZIP family, Mnt and Mga. Like Myc, the Mad proteins are tightly regulated with short half-lives. In general, Mad family members interfere with Myc-mediated processes, such as proliferation, transformation, and prevention of apoptosis by inhibiting transcription (3,4).

The Myc family is comprised of c-Myc, N-Myc, and L-Myc with often distinct patterns of expression during development as well as cancer (5). Amplification of each of the family members in cancer is frequently mutually exclusive with c-Myc being the most widely studied and most commonly amplified. N-Myc amplification, on the other hand, is found predominantly in neuroblastomas, and L-Myc amplification has been described in small cell lung cancer (SCLC) (6). Amplification of L-Myc in SCLC may result from chromosomal rearrangement between the rlf and MYCL genes (7). L-Myc expression by dendritic cells may be required for optimal T cell priming (8).

  1. Baudino, T.A. and Cleveland, J.L. (2001) Mol Cell Biol 21, 691-702.
  2. Blackwood, E.M. and Eisenman, R.N. (1991) Science 251, 1211-7.
  3. Henriksson, M. and Lüscher, B. (1996) Adv Cancer Res 68, 109-82.
  4. Grandori, C. et al. (2000) Annu Rev Cell Dev Biol 16, 653-99.
  5. Nesbit, C.E. et al. (1999) Oncogene 18, 3004-16.
  6. Nau, M.M. et al. Nature 318, 69-73.
  7. Mäkelä, T.P. et al. (1991) EMBO J 10, 1331-5.
  8. KC, W. et al. (2014) Nature 507, 243-7.

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 IP: Immunoprecipitation 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.
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 3
#76266

L-Myc (E3M5P) Rabbit mAb

Western Blotting Image 1: L-Myc (E3M5P) Rabbit mAb Expand Image
使用 L-Myc (E3M5P) Rabbit mAb(上图)、c-Myc (E5Q6W) Rabbit mAb #18583(上中图)、N-Myc (D4B2Y) Rabbit mAb #51705(下中图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对各种细胞系进行蛋白质印迹分析。每个 Myc 家族成员的细胞类型特异性是根据 RNAseq 数据预测的,并证实了抗体的特异性。NCI-H1963 细胞表达野生型和由染色体内重排产生的 RLF-L-Myc 融合蛋白。
Western Blotting Image 2: L-Myc (E3M5P) Rabbit mAb Expand Image
使用 L-Myc (E3M5P) Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对转染空载 (-) 或转染带有 Myc/DDK 标签的全长人 L-Myc 蛋白 (hMYCL-Myc/DDK; +) 表达载体的 293T 细胞提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: L-Myc (E3M5P) Rabbit mAb Expand Image
对 NCI-H209 细胞提取物 L-Myc 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 L-Myc (E3M5P) Rabbit mAb。使用 L-Myc (E3M5P) Rabbit mAb 进行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Chromatin Immunoprecipitation Image 1: L-Myc (E3M5P) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对来自 NCI-H209 细胞的交联染色质和 L-Myc (E3M5P) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析。使用 human ATF4 promoter primers、human NPM-1 intron 1 primers 和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 对富集的 DNA 进行定量分析。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。