Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-YTHDF2 antibody [EPR23544-19] - BSA and Azide free
See all YTHDF2 primary antibodies
Description

Rabbit monoclonal [EPR23544-19] to YTHDF2 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC	Human
IHC-P	Mouse
IP	Human
WB	Human

See all applications and species data

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: HeLa and NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysates.
General notes
ab275037 is the carrier-free version of ab246514. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm.

Maxpar^® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze.
Storage buffer

Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23544-19
Isotype

IgG
Research areas

Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilizedNIH/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 - Anti-YTHDF2 antibody [EPR23544-19] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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 (mouse embryonic fibroblast) 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] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)

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

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)

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

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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 - Anti-YTHDF2 antibody [EPR23544-19] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)

This data was developed using ab246514, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275037)