Revision 7

#86132Store 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, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q86U44

Entrez-Gene Id:

56339

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:200 - 1:800
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.

Specificity / Sensitivity

METTL3 (E3F2A) Rabbit mAb recognizes endogenous levels of total METTL3 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human METTL3 protein.

Background

Methyltransferase-like protein 3 (METTL3) and methyltransferase-like protein 14 (METTL14) are the two catalytic subunits of an N6-methyltransferase complex that methylates adenosine residues in RNA (1). Methylation of adenosine residues regulates mRNA splicing, processing, translation efficiency, editing and stability, in addition to regulating primary miRNA processing, and is critical for proper regulation of the circadian clock, embryonic stem cell self-renewal, immune tolerance, response to various stimuli, meiosis and mouse fertility (2,3). In this complex, METTL3 functions as the catalytic methyltransferase subunit and METTL14 functions as the target recognition subunit by binding to RNA (4). In addition, the Wilms' tumor 1-associating protein (WTAP) functions as a regulatory subunit and is required for accumulation of the complex to nuclear speckles, which are sites of RNA processing (5). Several studies suggest a role for this complex in cancer. METTL3 expression is elevated in lung adenocarcinoma where it promotes growth, survival, and invasion of human lung cancer cells (6). In addition, WTAP is overexpressed in a number of different cancers and positively regulates cell migration and invasion in glioblastoma and cholangiocarcinoma (7,8).

  1. Liu, J. et al. (2014) Nat Chem Biol 10, 93-5.
  2. Yue, Y. et al. (2015) Genes Dev 29, 1343-55.
  3. Wu, R. et al. (2016) Mol Biotechnol 58, 450-9.
  4. Wang, X. et al. (2016) Nature 534, 575-8.
  5. Ping, X.L. et al. (2014) Cell Res 24, 177-89.
  6. Lin, S. et al. (2016) Mol Cell 62, 335-45.
  7. Jin, D.I. et al. (2012) Cancer Sci 103, 2102-9.
  8. Jo, H.J. et al. (2013) J Gastroenterol 48, 1271-82.

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 IHC-P: Immunohistochemistry (Paraffin) 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 7
#86132

METTL3 (E3F2A) Rabbit mAb

Western Blotting Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) 兔单克隆抗体对不同细胞系的提取物进行蛋白印迹分析。
Western Blotting Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb #86132 对来自 COS-7 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: METTL3 (E3F2A) Rabbit mAb Expand Image
对来自 293T 细胞提取物的 METTL3 进行免疫沉淀 。泳道 1 为 10% 敲入,泳道 2 为兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900,泳道 3 为 METTL3 (E3F2A) 兔单克隆抗体。使用 METTL3 (E3F2A) 兔单克隆抗体和小鼠抗兔 IgG (特定构象) (L27A9) 单克隆抗体 (HRP 偶联物) #5127 进行蛋白印迹分析。
Immunohistochemistry Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的人结肠腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 2: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的人非小细胞肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的人大细胞神经内分泌肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 4: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的人 B 细胞非霍奇金氏淋巴瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 5: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的正常人肾脏进行免疫组织化学分析。
Immunohistochemistry Image 6: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的正常人胰腺进行免疫组织化学分析。
Immunohistochemistry Image 7: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的正常人心脏进行免疫组织化学分析。
Immunohistochemistry Image 8: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的正常小鼠脑进行免疫组织化学分析。
Immunohistochemistry Image 9: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb 对石蜡包埋的小鼠睾丸进行免疫组织化学分析。
Immunohistochemistry Image 10: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 METTL3 (E3F2A) Rabbit mAb(左图)或 METTL3 Rabbit mAb(右图)对石蜡包埋的人卵巢腺癌组织进行免疫组织化学分析。这两种抗体可检测 METTL3 上独特的非重叠表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 11: METTL3 (E3F2A) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 METTL3 (E3F2A) Rabbit mAb(左图)对石蜡包埋的人子宫内膜样腺癌细胞进行免疫组织化学分析。
Chromatin Immunoprecipitation Image 1: METTL3 (E3F2A) Rabbit mAb Expand Image
使用 SimpleChIP® 酶解染色质免疫共沉淀试剂盒(磁珠法) #9005 对 mES 细胞的交联染色质与 METTL3 (E3F2A) 兔单克隆抗体或正常兔 IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® Mouse Sox2 Exon1 Primers #30180、mouse KLF2 intron 2 primers 和 SimpleChIP® Mouse MYT-1 Promoter Primers #8985 进行实时 PCR 来对富集的 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。