Revision 3

#43123Store 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-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9BYJ9

Entrez-Gene Id:

54915

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200

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

YTHDF1 (E8R5L) Rabbit mAb recognizes endogenous levels of total YTHDF1 protein. Experiments utilizing overexpression constructs indicated that this antibody does not cross-react with other YTHDF proteins. Clone E9P6V is more sensitive than E8R5L by immunofluorescence.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human YTHDF1 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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation 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.
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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 3
#43123

YTHDF1 (E8R5L) Rabbit mAb

Western Blotting Image 1: YTHDF1 (E8R5L) Rabbit mAb Expand Image
Western blot analysis of extracts from HCT 116 cells, transfected with control siRNA (-) or YTHDF1 siRNA (+), using YTHDF1 (E8R5L) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: YTHDF1 (E8R5L) Rabbit mAb Expand Image
Western blot analysis of extracts from MCF7 and AGS cells using YTHDF1 (E8R5L) Rabbit mAb.
Immunoprecipitation Image 1: YTHDF1 (E8R5L) Rabbit mAb Expand Image
Immunoprecipitation of YTHDF1 protein from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is YTHDF1 (E8R5L) Rabbit mAb. Western blot analysis was performed using YTHDF1 (E8R5L) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as the secondary antibody.
Immunofluorescence Image 1: YTHDF1 (E8R5L) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HCT 116 cells, mock transfected (left, high-expressing) or transfected with YTHDF1 siRNA (right, low-expressing), using YTHDF1 (E8R5L) Rabbit mAb (green) and DAPI #4083 (blue).
Immunofluorescence Image 2: YTHDF1 (E8R5L) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of U-2 OS cells, untreated (left) or treated with sodium arsenite (right; 500 µM, 1 hr), using YTHDF1 (E8R5L) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).