Recombinant Anti-YTHDF2 antibody [EPR23544-19] - BSA and Azide free (ab275037)
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 - BSA and Azide free
- Suitable for: IP, ICC/IF, Flow Cyt (Intra), IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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
Suitable for: IP, ICC/IF, Flow Cyt (Intra), IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa and NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysates.
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General notes
ab275037 is the carrier-free version of ab246514.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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
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. Do Not Freeze. -
Storage buffer
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23544-19 -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab275037 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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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
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Notes |
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IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa). |
Target
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Sequence similarities
Contains 1 YTH domain. - Information by UniProt
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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
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Alternative names
- 9430020E02Rik antibody
- CAHL antibody
- CLL associated antigen KW 14 antibody
see all
Images
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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.
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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.
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This data was developed using ab246514, the same antibody clone in a different buffer formulation.
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.
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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.
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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.
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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 kDaThis 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.
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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 kDaThis 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. -
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.
-
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.
-
This data was developed using ab246514, the same antibody clone in a different buffer formulation.
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.
-
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.
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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Datasheet download
Certificate of Compliance
References (0)
ab275037 has not yet been referenced specifically in any publications.