Name: Anti-YTHDF3 antibody [EPR21912-3]
SKU: ab220161
Rating: 5 (3 reviews)

Rabbit Recombinant Monoclonal YTHDF3 antibody. Suitable for IP, WB, IHC-Fr, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 19 publications.

Images

Western blot - Anti-YTHDF3 antibody [EPR21912-3] (AB220161), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	WB	IHC-Fr	Flow Cyt (Intra)	IHC-P
Human	Not recommended	Tested	Tested	Expected	Tested	Tested
Mouse	Not recommended	Expected	Tested	Tested	Expected	Tested
Rat	Not recommended	Expected	Tested	Tested	Expected	Tested

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat, Mouse, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -
Species Rat	Dilution info 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/60	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Use at 1/500 (mouse, rat) and 1/100 (human) dilution. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Use at 1/500 (mouse, rat) and 1/100 (human) dilution. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Use at 1/500 (mouse, rat) and 1/100 (human) dilution. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

See full target information YTHDF3

Function

Specifically recognizes and binds N6-methyladenosine (m6A)-containing RNAs, and regulates their stability (PubMed:28106072, PubMed:28106076, PubMed:28281539, PubMed:32492408). M6A is a modification present at internal sites of mRNAs and some non-coding RNAs and plays a role in mRNA stability and processing (PubMed:22575960, PubMed:24284625, PubMed:28106072, PubMed:28281539, PubMed:32492408). Acts as a regulator of mRNA stability by promoting degradation of m6A-containing mRNAs via interaction with the CCR4-NOT complex or PAN3 (PubMed:32492408). The YTHDF paralogs (YTHDF1, YTHDF2 and YTHDF3) share m6A-containing mRNAs targets and act redundantly to mediate mRNA degradation and cellular differentiation (PubMed:28106072, PubMed:28106076, PubMed:32492408). Acts as a negative regulator of type I interferon response by down-regulating interferon-stimulated genes (ISGs) expression: acts by binding to FOXO3 mRNAs (By similarity). Binds to FOXO3 mRNAs independently of METTL3-mediated m6A modification (By similarity). Can also act as a regulator of mRNA stability in cooperation with YTHDF2 by binding to m6A-containing mRNA and promoting their degradation (PubMed:28106072). Recognizes and binds m6A-containing circular RNAs (circRNAs); circRNAs are generated through back-splicing of pre-mRNAs, a non-canonical splicing process promoted by dsRNA structures across circularizing exons (PubMed:28281539). Promotes formation of phase-separated membraneless compartments, such as P-bodies or stress granules, by undergoing liquid-liquid phase separation upon binding to mRNAs containing multiple m6A-modified residues: polymethylated mRNAs act as a multivalent scaffold for the binding of YTHDF proteins, juxtaposing their disordered regions and thereby leading to phase separation (PubMed:31292544, PubMed:31388144, PubMed:32451507). The resulting mRNA-YTHDF complexes then partition into different endogenous phase-separated membraneless compartments, such as P-bodies, stress granules or neuronal RNA granules (PubMed:31292544). May also recognize and bind N1-methyladenosine (m1A)-containing mRNAs: inhibits trophoblast invasion by binding to m1A-methylated transcripts of IGF1R, promoting their degradation (PubMed:32194978). Has some antiviral activity against HIV-1 virus: incorporated into HIV-1 particles in a nucleocapsid-dependent manner and reduces viral infectivity in the next cycle of infection (PubMed:32053707). May interfere with this early step of the viral life cycle by binding to N6-methyladenosine (m6A) modified sites on the HIV-1 RNA genome (PubMed:32053707).

Alternative names

YTH domain-containing family protein 3, DF3, YTHDF3

Recommended products

Rabbit Recombinant Monoclonal YTHDF3 antibody. Suitable for IP, WB, IHC-Fr, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 19 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR21912-3
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The YTH domain family protein 3 (YTHDF3) is an m6A reader protein known also as YTH3 or YTHDF2-like protein 3. Its molecular weight is approximately 64 kDa. YTHDF3 plays a role in the recognition of N6-methyladenosine (m6A)-modified RNA an important post-transcriptional modification. It commonly expresses in various tissues including the brain liver and kidney indicating its involvement in multiple cellular processes. YTHDF3 binds to m6A-modified mRNA influencing the stability and translation of these molecules and interacts with other members of the YTH family.

Biological function summary

YTHDF3 facilitates the process of mRNA metabolism by participating in the regulation of mRNA translation and degradation. It functions as part of a dynamic complex with YTHDF1 and YTHDF2 coordinating the fate of m6A-modified mRNAs. YTHDF3 supports mRNA decay in synergy with YTHDF2 and enhances translation efficiency in cooperation with YTHDF1. This involvement in regulating gene expression at the post-transcriptional level equips cells to respond rapidly to various stimuli aligning gene expression with cellular needs.

Pathways

YTHDF3 significantly impacts RNA processing and metabolism pathways. YTHDF3 together with YTHDF1 and YTHDF2 modulates the m6A-mediated mRNA decay pathway. It holds functional significance in the cellular response to stress and adaptations in energy metabolism. YTHDF3 interacts closely with proteins such as IGF2BP1 alongside the YTHDF proteins which are critical in maintaining RNA stability and translation thereby illustrating its role across significant cellular pathways.

Associated diseases and disorders

YTHDF3 has implications in cancer and neurodegenerative diseases. High YTHDF3 expression associates with various cancer types including gliomas and breast cancer where it can influence tumor growth and metastasis. In neurodegenerative diseases such as Alzheimer's disease YTHDF3 interacts with proteins like YTHDF1 and YTHDF2 affecting RNA metabolism in neurons contributing potentially to the pathology of these disorders. Through these interactions YTHDF3 could impact disease progression and outcomes making it a target of interest in therapeutic research.

Product promise

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11 product images

Western blot - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
The wild-type and YTHDF3 knockout cell lysates were kindly provided by an anonymous collaborator.
ab220161 was shown to specifically react with YTHDF3 in wild-type mESC cells as signal was lost in YTHDF3 knockout cells. Wild-type and YTHDF3 knockout samples were subjected to SDS-PAGE. ab220161 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.
The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument using the ECL technique.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-YTHDF3 antibody [EPR21912-3] (ab220161) at 1/1000 dilution
Lane 1: Wild-type mESC (mouse embryo stem cell) whole cell lysate at 20 µg
Lane 2: YTHDF3 knockout mESC whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 64 kDa
Observed band size: 73 kDa
Exposure time: 59s
Immunoprecipitation - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
YTHDF3 was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab220161 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220161 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab220161 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab220161 in HeLa whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 15 seconds.
Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.
All lanes: Immunoprecipitation - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Predicted band size: 64 kDa
Observed band size: 73 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling YTHDF3 with ab220161 at 1/100 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human colon (PMID:29103884, 28250115) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Frozen sections) - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling YTHDF3 with ab220161 at 1/100 dilution (green), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Cytoplasmic and nuclear staining in mouse cerebrum (PMID:29103884, 28250115) is observed. Counterstained with DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1,000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).
Flow Cytometry (Intracellular) - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling YTHDF3 with ab220161 at 1/60 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Lysate should be made freshly and used in WB immediately to minimize protein degradation (lane1-4). Lane 5-7 are the lysates from same cell lines but have experienced freeze-thaw cycles.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-4: 37 secs; Lanes 5-7: 92 secs.
All lanes: Western blot - Anti-YTHDF3 antibody [EPR21912-3] (ab220161) at 1/1000 dilution
Lane 1: HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate (fresh lysate) at 20 µg
Lane 2: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate (fresh lysate) at 20 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate (fresh lysate) at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate (fresh lysate) at 20 µg
Lane 5: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 6: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 7: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 64 kDa
Western blot - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
The YTHDF recombinant proteins were kindly provided by an anonymous collaborator.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-YTHDF3 antibody [EPR21912-3] (ab220161) at 1/5000 dilution
Lane 1: GST-tagged human YTHDF1 recombinant protein 20 ng
Lane 2: GST-tagged human YTHDF2 recombinant protein 20 ng
Lane 3: GST-tagged human YTHDF3 recombinant protein 20 ng
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 64 kDa
Exposure time: 10s
Immunohistochemistry (Frozen sections) - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum tissue labeling YTHDF3 with ab220161 at 1/100 dilution (green), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Cytoplasmic and nuclear staining in rat cerebrum (PMID:29103884, 28250115) is observed. Counterstained with DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1,000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling YTHDF3 with ab220161 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on neurons of rat cerebrum (PMID:29103884, 28250115) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling YTHDF3 with ab220161 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on neurons of mouse cerebrum (PMID:29103884, 28250115) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
This image is courtesy of a customer review submitted by Bin Shen
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF3 antibody [EPR21912-3] (ab220161)
Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde-fixed mouse ovary tissue permeabilized with PBST (0.5% Triton X-100 in PBS). Stained with ab220161 at 1/200 dilution. Secondary antibody used was Alex Fluor 555 donkey anti-rabbit IgG at 1/500 dilution. Blocking was done with 5% BSA in PBST (0.1% tween in PBS) for 1 hour 30 minutes at 25°C. The sample was incubated with the primary antibody and 5% BSA in PBST (0.1% tween in PBS) for 12 hours at 4°C. Antigen retrieval method was heat mediated, Citrate antigen repair solution.