Revision 3

#71283Store 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, IF-F, IF-IC

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

65

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9Y5A9

Entrez-Gene Id:

51441

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Frozen) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200

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

YTHDF2 (E2I2H) Rabbit mAb recognizes endogenous levels of total YTHDF2 protein. Experiments utilizing overexpression constructs indicated that this antibody does not cross-react with other YTHDF proteins.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly167 of human YTHDF2 protein.

Background

N6-methyladenosine (m6 A) is an abundant RNA modification that plays an important role in mRNA splicing, processing, and stability. The m6 A modification is specifically recognized by members of the YT521B homology (YTH) domain-containing family (YTHDF), consisting of YTHDF1, YTHDF2, and YTHDF3. All three members of the YTHDF family are primarily cytosolic proteins that share similar sequence and domain structure, including a conserved C-terminal YTH domain that specifically interacts with m6 A (1). Despite these similarities, recent studies suggest that YTHDF proteins are involved in distinct regulatory functions with minimal overlap. Specifically, YTHDF1 binding has been reported to promote enhanced mRNA translation, but has no measurable effect on mRNA stability (2). Conversely, YTHDF2 binding appears to promote mRNA degradation, but has minimal effect on translation efficiency (3). The function of YTHDF3 is less clear, but it has been proposed to function as an auxiliary protein for both YTHDF1 and YTHDF2, helping to promote either increased mRNA translation or decay, respectively (4). Additional studies offer a different viewpoint, suggesting that all three YTHDF proteins initiate mRNA degradation (5), or mediate increased mRNA stability and protein expression (6), promoting the idea that these proteins may carry out similar rather than distinct functions.

  1. Meyer, K.D. and Jaffrey, S.R. (2017) Annu Rev Cell Dev Biol 33, 319-42.
  2. Wang, X. et al. (2015) Cell 161, 1388-99.
  3. Wang, X. et al. (2014) Nature 505, 117-20.
  4. Shi, H. et al. (2017) Cell Res 27, 315-28.
  5. Du, H. et al. (2016) Nat Commun 7, 12626.
  6. Kennedy, E.M. et al. (2016) Cell Host Microbe 19, 675-85.

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 IP: Immunoprecipitation 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 3
#71283

YTHDF2 (E2I2H) Rabbit mAb

Western Blotting Image 1: YTHDF2 (E2I2H) Rabbit mAb Expand Image
使用 YTHDF2 (E2I2H) Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对转染非靶向 siRNA (siNT) 或靶向 YTHDF2 (siYTHDF2) 的 siRNA 的 293T 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 2: YTHDF2 (E2I2H) Rabbit mAb Expand Image
使用 YTHDF2 (E2I2H) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: YTHDF2 (E2I2H) Rabbit mAb Expand Image
对 293T 细胞提取物 YTHDF2 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 YTHDF2 (E2I2H) Rabbit mAb。使用 YTHDF2 (E2I2H) 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。
Immunofluorescence Image 1: YTHDF2 (E2I2H) Rabbit mAb Expand Image
使用 YTHDF2 (E2I2H) Rabbit mAb(红色)和 GFAP (GA5) Mouse mAb (Alexa Fluor® 488 Conjugate) #3655(绿色)对小鼠海马进行共聚焦免疫荧光分析。肌动蛋白丝用 DyLight 650 Phalloidin #12956(伪彩青色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Immunofluorescence Image 2: YTHDF2 (E2I2H) Rabbit mAb Expand Image
使用 YTHDF2 (E2I2H) Rabbit mAb(红色)和 GFAP (GA5) Mouse mAb (Alexa Fluor® 488 Conjugate) #3655(绿色)对小鼠延髓进行共聚焦免疫荧光分析。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Immunofluorescence Image 3: YTHDF2 (E2I2H) Rabbit mAb Expand Image
使用 YTHDF2 (E2I2H) Rabbit mAb(红色)和 GFAP (GA5) Mouse mAb (Alexa Fluor® 488 Conjugate) #3655(绿色)对小鼠视网膜进行共聚焦免疫荧光分析。肌动蛋白丝用 DyLight 650 Phalloidin #12956(伪彩青色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Immunofluorescence Image 1: YTHDF2 (E2I2H) Rabbit mAb Expand Image
使用 YTHDF2 (E2I2H) Rabbit mAb(绿色)对转染空载(左图,高表达)或转染针对人 YTHDF2 的 siRNA(右图,低表达)的 HCT 116 细胞进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Immunofluorescence Image 2: YTHDF2 (E2I2H) Rabbit mAb Expand Image
使用 YTHDF2 (E2I2H) Rabbit mAb(绿色)和 G3BP1 (TT-Y) Mouse mAb(红色)对未经处理(左图)或经过亚砷酸钠(500 µM,1 小时)(右图)处理的 U-2 OS 细胞进行共聚焦免疫荧光分析。在用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离二抗结合位点后,随后用 GM130 (D6B1) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #59890(伪彩青色)标记该细胞。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。