Revision 1

#80303Store 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):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q7Z739

Entrez-Gene Id:

253943

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Frozen) 1:400
Immunofluorescence (Immunocytochemistry) 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

YTHDF3 (E2J9I) Rabbit mAb recognizes endogenous levels of total YTHDF3 protein. This antibody does not cross-react with YTHDF1 or YTHDF2 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly169 of human YTHDF3 protein.

Background

N6-methyladenosine (m6A) is an abundant RNA modification that plays an important role in mRNA splicing, processing, and stability. The m6A modification is specifically recognized by YT521B homology (YTH) domain-containing proteins, consisting of five members in mammals: YTH domain-containing proteins 1 and 2 (YTHDC1 and YTHDC2), and YTH domain-containing family proteins 1, 2, and 3 (YTHDF1, YTHDF2, and YTHDF3) (1). YTHDC1, also known as splicing factor YT521, regulates alternative splicing by functioning as a key regulator of exon-inclusion or exon-skipping. YTHDC1 promotes exon-inclusion by recruiting pre-mRNA splicing factor SRSF3 to regions containing m6A, while repressing exon-skipping by blocking SRSF10 binding to these same regions (2). Increased expression of YTHDC1 promotes malignant endometrial carcinoma (EC) through alternative splicing of vascular endothelial growth factor-A (VEGF-A), resulting in an increase in VEGF-165 isoform and increased EC cell invasion (3). YTHDC2 functions to enhance the translation efficiency of target mRNAs and may play a role in spermatogenesis (4). All three members of the YTHDF family are paralogs that share similar sequence and domain structure, including the conserved C-terminal YTH domain that specifically interacts with m6A (5). 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 (6). Conversely, YTHDF2 binding appears to promote mRNA degradation, but has minimal effect on translation efficiency (7). 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 (8). Additional studies offer a different viewpoint, suggesting that all three YTHDF proteins initiate mRNA degradation, or mediate increased mRNA stability and protein expression, promoting the idea that these proteins may carry out similar rather than distinct functions (9,10).

  1. Xu, C. et al. (2015) J Biol Chem 290, 24902-13.
  2. Xiao, W. et al. (2016) Mol Cell 61, 507-519.
  3. Zhang, B. et al. (2015) Tumour Biol 37, 15543-9.
  4. Hsu, P.J. et al. (2017) Cell Res 27, 1115-1127.
  5. Meyer, K.D. and Jaffrey, S.R. (2017) Annu Rev Cell Dev Biol 33, 319-342.
  6. Wang, X. et al. (2015) Cell 161, 1388-99.
  7. Wang, X. et al. (2014) Nature 505, 117-20.
  8. Shi, H. et al. (2017) Cell Res 27, 315-328.
  9. Du, H. et al. (2016) Nat Commun 7, 12626.
  10. 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 BSA, 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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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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
#80303

YTHDF3 (E2J9I) Rabbit mAb

Western Blotting Image 1: YTHDF3 (E2J9I) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using YTHDF3 (E2J9I) Rabbit mAb.
Western Blotting Image 2: YTHDF3 (E2J9I) Rabbit mAb Expand Image
Western blot analysis of extracts from HCT 116 cells, transfected with control siRNA (-) or YTHDF3 siRNA (+), using YTHDF3 (E2J9I) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: YTHDF3 (E2J9I) Rabbit mAb Expand Image
对 293T 细胞提取物的 YTHDF3 蛋白进行免疫沉淀分析。Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is YTHDF3 (E2J9I) Rabbit mAb. Western blot analysis was performed using YTHDF3 (E2J9I) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunofluorescence Image 1: YTHDF3 (E2J9I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse colon using YTHDF3 (E2J9I) Rabbit mAb (left, green), DyLight 554 Phalloidin #13054 (right, red), and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 2: YTHDF3 (E2J9I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum using YTHDF3 (E2J9I) Rabbit mAb (left, green), GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657 (right, red), and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 3: YTHDF3 (E2J9I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse hippocampus using YTHDF3 (E2J9I) Rabbit mAb (left, green), GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657 (right, red), and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 1: YTHDF3 (E2J9I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HCT 116 cells, mock transfected (left, high-expressing) or transfected with siRNA directed against human YTHDF3 (right, low-expressing), using YTHDF3 (E2J9I) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
Immunofluorescence Image 2: YTHDF3 (E2J9I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with sodium arsenite (500 µM, 1 hr; right), using YTHDF3 (E2J9I) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).