Revision 1

#52901Store 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, IHC-P, IF-F, IF-IC

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

100

Source/Isotype:

Rabbit IgG

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Frozen) 1:1600 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:3200 - 1:12800

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

NSUN2 (E9Y5E) Rabbit mAb recognizes endogenous levels of total NSUN2 protein. Species reactivity for IHC-P is human only.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human NSUN2 protein.

Background

Chemical modifications of RNA regulate many cellular processes. One particular RNA modification, 5-methylcytosine (5-mC), regulates ribosome assembly, translation, and RNA stability (1). In eukaryotes, this modification is added to RNA by the DNA methyltransferase homologue 2 protein (DNMT2; also known as TRDMT1), and also by members of the NOL1/NOP2/SUN domain (NSUN) family of proteins. NSUN proteins are putative S-adenosylmethionine (SAM)-dependent methyltransferases that carry out their enzymatic activity by utilizing two cysteine residues in their active sites (2). There are currently seven known members of this family, consisting of NOP2 (NSUN1) and NSUN2-7.

NSUN2 is an 86 kDa member of the NSUN family that is predominantly localized to the nucleus and methylates a variety of tRNAs and other RNA substrates (3). NSUN2 has been found to play a role in the methylation of pre-tRNALeu, tRNAGly, tRNAVal, tRNALeu, and tRNAAsp (1,4,5). It has also been found to methylate mRNAs and several ncRNAs, such as vtRNAs (6-8). Elevated NSUN2 protein levels have been reported in multiple types of human cancers, including breast, prostate, kidney, bladder, and liver, among others (9). The methyltransferase activity of NSUN2 is reduced upon Aurora B kinase-mediated phosphorylation at Ser139 (10), and there is potential for NSUN2 to be used as a therapeutic target or diagnostic marker for Aurora B linked cancers (9).

  1. Trixl, L. and Lusser, A. (2019) Wiley Interdiscip Rev RNA 10, e1510.
  2. Bohnsack, K.E. et al. (2019) Genes (Basel) 10, pii: E102. doi: 10.3390/genes10020102.
  3. Bourgeois, G. et al. (2015) PLoS One 10, e0133321.
  4. Brzezicha, B. et al. (2006) Nucleic Acids Res 34, 6034-43.
  5. Tuorto, F. et al. (2012) Nat Struct Mol Biol 19, 900-5.
  6. Tang, H. et al. (2015) Aging (Albany NY) 7, 1143-58.
  7. Xing, J. et al. (2015) Mol Cell Biol 35, 4043-52.
  8. Hussain, S. et al. (2013) Cell Rep 4, 255-61.
  9. Okamoto, M. et al. (2012) DNA Cell Biol 31, 660-71.
  10. Sakita-Suto, S. et al. (2007) Mol Biol Cell 18, 1107-17.

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 IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

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 1
#52901

NSUN2 (E9Y5E) Rabbit mAb

Western Blotting Image 1: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb(上图)或 α-Actinin (D6F6) XP® Rabbit mAb #6487(下图),对经对照 siRNA(-)或 NSUN2 siRNA(+)转染的 293T 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 2: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb 对 HeLa 细胞、HT-1080 细胞和 Neuro-2a 细胞的提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) 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: NSUN2 (E9Y5E) Rabbit mAb Expand Image

使用 NSUN2 (E9Y5E) Rabbit mAb 对石蜡包埋的人胃肠道间质瘤组织进行免疫组织化学分析。

Immunohistochemistry Image 3: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb 对石蜡包埋的人前列腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 4: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb 对石蜡包埋的人胃腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 5: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb 对石蜡包埋的人正常胎盘组织进行免疫组织化学分析。
Immunohistochemistry Image 6: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb(上图)或 NSUN2 Rabbit mAb(下图)对石蜡包埋的人结肠癌组织(左图)、鳞状细胞癌组织(中图)或食道癌组织(右图)进行免疫组织化学分析。这两种抗体可检测人 NSUN2 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 7: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb 对石蜡包埋的人正常皮肤组织进行免疫组织化学分析。
Immunohistochemistry Image 8: NSUN2 (E9Y5E) Rabbit mAb Expand Image
与浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)比较,使用 NSUN2 (E9Y5E) Rabbit mAb(左图)对石蜡包埋的人阑尾组织进行免疫组织化学分析。
Immunofluorescence Image 1: NSUN2 (E9Y5E) Rabbit mAb Expand Image
对使用 NSUN2 (E9Y5E) Rabbit mAb(左图,绿色)标记的固定冷冻的小鼠结肠组织进行共聚焦免疫荧光分析。然后使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离的二抗结合位点,之后使用 Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182(右图,红色)和 DAPI #4083(右图,蓝色)进行共同标记。
Immunofluorescence Image 2: NSUN2 (E9Y5E) Rabbit mAb Expand Image
对使用 NSUN2 (E9Y5E) Rabbit mAb(左图,绿色)标记的固定冷冻的小鼠睾丸组织进行共聚焦免疫荧光分析。然后使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离的二抗结合位点,之后使用 Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182(右图,红色)和 DAPI #4083(右图,蓝色)进行共同标记。
Immunofluorescence Image 3: NSUN2 (E9Y5E) Rabbit mAb Expand Image
对使用 NSUN2 (E9Y5E) Rabbit mAb(左图,绿色)标记的固定冷冻的小鼠肝脏组织进行共聚焦免疫荧光分析。然后使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离的二抗结合位点,之后使用 Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182(右图,红色)和 DAPI #4083(右图,蓝色)进行共同标记。
Immunofluorescence Image 4: NSUN2 (E9Y5E) Rabbit mAb Expand Image
对使用 NSUN2 (E9Y5E) Rabbit mAb(左图,绿色)标记的固定冷冻的小鼠胰腺组织进行共聚焦免疫荧光分析。然后使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离的二抗结合位点,之后使用 Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182(右图,红色)和 DAPI #4083(右图,蓝色)进行共同标记。
Immunofluorescence Image 1: NSUN2 (E9Y5E) Rabbit mAb Expand Image
使用 NSUN2 (E9Y5E) Rabbit mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色),对经对照 siRNA(左图)或 NSUN2 siRNA(右图)转染的 HCT 116 细胞进行共聚焦免疫荧光分析。