WB, IP, IF-F, IF-IC
H M R Mk
Endogenous
70
Rabbit IgG
#Q7Z739
253943
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:100 |
| Immunofluorescence (Frozen) | 1:400 |
| Immunofluorescence (Immunocytochemistry) | 1:3200 |
Storage
Specificity / Sensitivity
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).
- Xu, C. et al. (2015) J Biol Chem 290, 24902-13.
- Xiao, W. et al. (2016) Mol Cell 61, 507-519.
- Zhang, B. et al. (2015) Tumour Biol 37, 15543-9.
- Hsu, P.J. et al. (2017) Cell Res 27, 1115-1127.
- Meyer, K.D. and Jaffrey, S.R. (2017) Annu Rev Cell Dev Biol 33, 319-342.
- Wang, X. et al. (2015) Cell 161, 1388-99.
- Wang, X. et al. (2014) Nature 505, 117-20.
- Shi, H. et al. (2017) Cell Res 27, 315-328.
- Du, H. et al. (2016) Nat Commun 7, 12626.
- 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
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