Recombinant Anti-YTHDF1 + YTHDF3 + YTHDF2 antibody [EPR26183-69] (ab290734)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26183-69] to YTHDF1 + YTHDF3 + YTHDF2
- Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-YTHDF1 + YTHDF3 + YTHDF2 antibody [EPR26183-69] -
Description
Rabbit monoclonal [EPR26183-69] to YTHDF1 + YTHDF3 + YTHDF2 -
Host species
Rabbit -
Specificity
Please note that this antibody does not react with Rat species for ICC and Flow Cyt (intra) applications.
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Tested applications
Suitable for: WB, IP, Flow Cyt (Intra), ICC/IFmore details
Unsuitable for: 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: HeLa (human cervix adenocarcinoma epithelial cell) and 293T (human embryonic kidney epithelial cell) whole cell lysates. NIH/3T3 (mouse embryonic fibroblast) whole cell lysate. PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate. ICC/IF: HeLa cells, NIH/3T3 cells. Flow Cyt (intra): HeLa cells, NIH/3T3 cells. IP: HeLa and NIH/3T3 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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR26183-69 -
Isotype
IgG -
Research areas
Associated products
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab290734 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. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
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IP |
1/30.
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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/50.
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Notes |
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WB
1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa). |
IP
1/30. |
Flow Cyt (Intra)
1/500. |
ICC/IF
1/50. |
Images
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All lanes : Anti-YTHDF1 + YTHDF3 + YTHDF2 antibody [EPR26183-69] (ab290734) at 1/1000 dilution
Lane 1 : HeLa (human cervix 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) whole cell lysate.
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 61 kDa
Observed band size: 61 kDa
Exposure time: 70 secondsBlocking / Diluent buffer and concentration:5% NFDM/TBST.
Lysates should be made freshly and used in WB immediately to minimize protein degradation.
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All lanes : Anti-YTHDF1 + YTHDF3 + YTHDF2 antibody [EPR26183-69] (ab290734) at 1/1000 dilution
Lane 1 : His-tagged human YTHDF1 recombinant protein 10ng
Lane 2 : His-tagged human YTHDF2 recombinant protein 10ng
Lane 3 : His-tagged human YTHDF3 recombinant protein 10ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 61 kDa
Observed band size: 61 kDa
Exposure time: 70 secondsBlocking / Diluting buffer and concentration: 5% NFDM/TBST
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized NIH/3T3 (mouse embryonic fibroblast) cells lebelling YTHDF1 + YTHDF2 + YTHDF3 with ab290734 at 1/50 (10.04 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/mL) dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/mL) dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized HeLa (human cervix adenocarcinoma epithelial cell) cells lebelling YTHDF1 + YTHDF2 + YTHDF3 with ab290734 at 1/50 (10.04 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/mL) dilution (Green). Confocal image showing mostly cytoplasmic staining in Hela cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/mL) dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling YTHDF1 + YTHDF2 + YTHDF3 with ab290734 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, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling YTHDF1 + YTHDF2 + YTHDF3 with ab290734 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, ab150081) at 1/2000 dilution was used as the secondary antibody.
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YTHDF1 + YTHDF2 + YTHDF3 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug with ab290734 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab290734 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug
Lane 2: ab290734 IP in NIH/3T3 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab290734 in NIH/3T3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds
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YTHDF1 + YTHDF2 + YTHDF3 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug ab290734 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using anti YTHDF1 + YTHDF2 + YTHDF3 antibody (ab290734) at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg
Lane 2: anti YTHDF1 + YTHDF2 + YTHDF3 antibody (ab290734) IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab290734 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds
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 (1)
ab290734 has been referenced in 1 publication.
- Lin W et al. Tumor-intrinsic YTHDF1 drives immune evasion and resistance to immune checkpoint inhibitors via promoting MHC-I degradation. Nat Commun 14:265 (2023). PubMed: 36650153