Name: Anti-YTHDF2 antibody [EPR23544-19]
Brand: RabMAb
SKU: ab246514
Rating: 4 (3 reviews)

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Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23544-19] to YTHDF2
Suitable for: WB, IHC-P, Flow Cyt (Intra), IP, ICC
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-YTHDF2 antibody [EPR23544-19]
See all YTHDF2 primary antibodies
Description

Rabbit monoclonal [EPR23544-19] to YTHDF2
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, Flow Cyt (Intra), IP, ICCmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, LNCaP, MEF and PC-12 whole cell lysates; His-tagged human YTHDF2 recombinant protein. IHC-P: Human testis tissue; Mouse testis tissue; Rat testis tissue. ICC: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa and NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysates.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23544-19
Isotype

IgG
Research areas

Alternative Versions
- Anti-YTHDF2 antibody [EPR23544-19] - BSA and Azide free (ab275037)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab246514 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB	(1)	1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra)		1/500.
IP		1/30.
ICC		1/50.

Notes
WB 1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra) 1/500.
IP 1/30.
ICC 1/50.

Sequence similarities

Contains 1 YTH domain.
Target information above from: UniProt accession Q9Y5A9 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 51441 Human
- Entrez Gene: 213541 Mouse
- Entrez Gene: 313053 Rat
- Omim: 610640 Human
- SwissProt: Q9Y5A9 Human
- SwissProt: Q91YT7 Mouse
- Unigene: 532286 Human
Alternative names
- 9430020E02Rik antibody
- CAHL antibody
- CLL associated antigen KW 14 antibody
- CLL-associated antigen KW-14 antibody
- HGRG8 antibody
- High glucose regulated protein 8 antibody
- High-glucose-regulated protein 8 antibody
- NY REN 2 antibody
- Renal carcinoma antigen NY REN 2 antibody
- Renal carcinoma antigen NY-REN-2 antibody
- YTH domain family 2 antibody
- YTH domain family member 2 antibody
- YTH domain family protein 2 antibody
- YTH N6 methyladenosine RNA binding protein 2 antibody
- YTHD2_HUMAN antibody
- YTHDF 2 antibody
- YTHDF2 antibody
see all

Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial adenocarcinoma cell line) (siRNA control) and HeLa (siRNA YTHDF) cells labelling YTHDF2 with ab246514 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line transfected with scramble siRNA and reduced staining in HeLa cell line transfected with siRNA YTHDF2. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryo fibroblast cell line) cells labelling YTHDF2 with ab246514 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling YTHDF2 with ab246514 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

YTHDF2 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate with ab246514 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246514 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: NIH/3T3 whole cell lysate 10 ug

Lane 2: ab246514 IP in NIH/3T3 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246514 in NIH/3T3 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 84 seconds.

Lysates were made freshly and used in IP test immediately to minimize protein degradation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling YTHDF2 with ab246514 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in mouse testis (PMID: 30903744, PMID: 29103884). The section was incubated with ab246514 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

All lanes : Anti-YTHDF2 antibody [EPR23544-19] (ab246514) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : LNCaP (human prostate carcinoma epithelial cell) whole cell lysate
Lane 3 : MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 62 kDa
Observed band size: 62 kDa

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-2: 37 seconds; Lane 3: 180 seconds; Lane 4:37 seconds.

The molecular mass observed is consistent with the literature (PMID: 28104805).

Lysates should be made freshly and used in WB immediately to minimize protein degradation.
Western blot - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

All lanes : Anti-YTHDF2 antibody [EPR23544-19] (ab246514) at 1/5000 dilution

Lane 1 : LIF-His-tagged human YTHDF1 recombinant protein, 10 ng
Lane 2 : His-tagged human YTHDF2 recombinant protein, 10 ng
Lane 3 : His-tagged human YTHDF3 recombinant protein, 10 ng

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 62 kDa
Observed band size: 62 kDa

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 59 seconds.

These rec proteins were made in house.
LIF-His-tagged human YTHDF1 recombinant protein was expressed from a mammalian - HEK-293 expression system.
His-tagged human YTHDF2 recombinant protein is extracts of E.coli expressing YTHDF2.
His-tagged human YTHDF3 recombinant protein is extracts of E.coli expressing YTHDF3.
Immunoprecipitation - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

YTHDF2 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell line) whole cell lysate with ab246514 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246514 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HeLa whole cell lysate 10 ug

Lane 2: ab246514 IP in HeLa whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246514 in HeLa whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling YTHDF2 with ab246514 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in rat testis (PMID: 30903744, PMID: 29103884). The section was incubated with ab246514 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry (Intracellular) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labelling YTHDF2 with ab246514 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling YTHDF2 with ab246514 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human testis (PMID: 30903744, PMID: 29103884).The section was incubated with ab246514 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab246514? Please let us know so that we can cite the reference in this datasheet.

ab246514 has been referenced in 2 publications.

Xu H et al. YTHDF2 alleviates cardiac hypertrophy via regulating Myh7 mRNA decoy. Cell Biosci 11:132 (2021). PubMed: 34266473
Shen X et al. YTHDF2 Inhibits Gastric Cancer Cell Growth by Regulating FOXC2 Signaling Pathway. Front Genet 11:592042 (2020). PubMed: 33505426

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1-3 of 3 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Zebrafish Cell (whole embryo 3.5-4.5 hours past fertilization)

Permeabilization

Yes - -20°C MeOH storage over night

Specification

whole embryo 3.5-4.5 hours past fertilization

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 23°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 14 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (T cells)

Gel Running Conditions

Reduced Denaturing

Loading amount

15 µg

Specification

T cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 09 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Pig Cell (Swine Testicle (ST) cells)

Permeabilization

Yes - Triton-X 100

Specification

Swine Testicle (ST) cells

Blocking step

Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 22°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Oct 12 2020

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Sequence similarities

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-YTHDF2 antibody [EPR23544-19]

Western blot abreview for Anti-YTHDF2 antibody [EPR23544-19]

Immunocytochemistry/ Immunofluorescence abreview for Anti-YTHDF2 antibody [EPR23544-19]