Recombinant Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22349-41] to YTHDF1
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-YTHDF1 antibody [EPR22349-41]
See all YTHDF1 primary antibodies -
Description
Rabbit monoclonal [EPR22349-41] to YTHDF1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: GST-tagged human YTHDF1 recombinant protein. Wild-type mESC whole cell lysate. HepG2, NIH/3T3 and HeLa whole cell lysate. Human brain lysate. Rat testis lysate. IP: YTHDF1 IP in HepG2 whole cell lysate.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22349-41 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab220162 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 |
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WB |
1/1000. Predicted molecular weight: 61 kDa.
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IP |
1/30.
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Notes |
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WB
1/1000. Predicted molecular weight: 61 kDa. |
IP
1/30. |
Target
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Sequence similarities
Contains 1 YTH domain. - Information by UniProt
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Database links
- Entrez Gene: 54915 Human
- Entrez Gene: 228994 Mouse
- Entrez Gene: 296467 Rat
- SwissProt: Q9BYJ9 Human
- SwissProt: P59326 Mouse
- Unigene: 11747 Human
- Unigene: 30205 Mouse
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Alternative names
- C20orf21 antibody
- DACA 1 antibody
- DACA-1 antibody
see all
Images
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All lanes : Anti-YTHDF1 antibody [EPR22349-41] (ab220162) at 1/1000 dilution
Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsLysates were freshly made and used for Western blotting immediately to minimize protein degradation.
Blocking and diluting buffer and concntration: 5% NFDM/TBST
Imaging carried out using iBright CL 1000 Imaging System -
YTHDF1 was immunoprecipitated from HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab220162 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220162 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab220162 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220162 in HeLa whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.Lysate was freshly made and used for Western blotting immediately to minimize protein degradation.
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All lanes : Anti-YTHDF1 antibody [EPR22349-41] (ab220162) at 1/1000 dilution
Lane 1 : Wild-type mESC (mouse embryo stem cell) whole cell lysate
Lane 2 : YTHDF1 knockout mESC whole cell lysate
Lane 3 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 61 kDa
Observed band size: 61 kDaThe wild-type and YTHDF1 knockout cell lysates were kindly provided by an anonymous collaborator.
ab220162 was shown to specifically react with YTHDF1 in wild-type mESC cells as signal was lost in YTHDF1 knockout cells. Wild-type and YTHDF1 knockout samples were subjected to SDS-PAGE. ab220162 and 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 (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.
Exposure times: Lanes 1 and 2: 59 seconds; Lanes 3 and 4: 26 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-YTHDF1 antibody [EPR22349-41] (ab220162) at 1/1000 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 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 61 kDa
Observed band size: 87 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsThe YTHDF recombinant proteins were kindly provided by an anonymous collaborator.
Blocking/Dilution buffer: 5% NFDM/TBST.
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YTHDF1 was immunoprecipitated from 0.35 mg HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate with ab220162 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220162 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HepG2 whole cell lysate 10 µg (Input).
Lane 2: ab220162 IP in HepG2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220162 in HepG2 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 seconds.Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (14)
ab220162 has been referenced in 14 publications.
- Wei K et al. Methylation recognition protein YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) regulates the proliferation, migration and invasion of osteosarcoma by regulating m6A level of CCR4-NOT transcription complex subunit 7 (CNOT7). Bioengineered 13:5236-5250 (2022). WB ; Human . PubMed: 35156522
- You A et al. TTC22 promotes m6A-mediated WTAP expression and colon cancer metastasis in an RPL4 binding-dependent pattern. Oncogene N/A:N/A (2022). WB ; Human . PubMed: 35798874
- Jiang Z et al. Circular RNA protein tyrosine kinase 2 (circPTK2) promotes colorectal cancer proliferation, migration, invasion and chemoresistance. Bioengineered 13:810-823 (2022). PubMed: 34974791
- Wang Y et al. HLA complex P5 upregulation is correlated with poor prognosis and tumor progression in esophageal squamous cell carcinoma. Bioengineered 13:9301-9311 (2022). PubMed: 35389828
- Li P et al. ELK1-mediated YTHDF1 drives prostate cancer progression by facilitating the translation of Polo-like kinase 1 in an m6A dependent manner. Int J Biol Sci 18:6145-6162 (2022). PubMed: 36439881