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Knockout Tested Rabbit Recombinant Monoclonal WTAP phospho S341 antibody. Carrier free. Suitable for WB, IHC-P, IP, Dot and reacts with Human, Synthetic peptide samples.

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Images

Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (AB309351), expandable thumbnail
  • Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (AB309351), expandable thumbnail
  • Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (AB309351), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (AB309351), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (AB309351), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PIPDotICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Expected
Not recommended
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Synthetic peptide
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Synthetic peptide

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Synthetic peptide

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Synthetic peptide

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat, Synthetic peptide

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat, Synthetic peptide

Dilution info

-

Notes

-

Target data

Function

Associated component of the WMM complex, a complex that mediates N6-methyladenosine (m6A) methylation of RNAs, a modification that plays a role in the efficiency of mRNA splicing and RNA processing (PubMed:29507755). Required for accumulation of METTL3 and METTL14 to nuclear speckle (PubMed:24316715, PubMed:24407421, PubMed:24981863). Acts as a mRNA splicing regulator (PubMed:12444081). Regulates G2/M cell-cycle transition by binding to the 3' UTR of CCNA2, which enhances its stability (PubMed:17088532). Impairs WT1 DNA-binding ability and inhibits expression of WT1 target genes (PubMed:17095724).

Alternative names

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal WTAP phospho S341 antibody. Carrier free. Suitable for WB, IHC-P, IP, Dot and reacts with Human, Synthetic peptide samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR27039-57

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

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.

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.

Supplementary info

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

Activity summary

WTAP also known as Wilms tumor 1-associated protein is a core component of the writer complex that catalyzes N6-methyladenosine (m6A) methylation on mRNA. With a molecular mass of approximately 39 kDa WTAP localizes almost ubiquitously in human tissues but shows higher expression in liver and kidney cells. This protein assists in the regulation of mRNA splicing and stability functioning as an adapter for RNA binding proteins.

Biological function summary

This protein plays a critical role in post-transcriptional gene regulation by being part of the m6A methyltransferase complex which includes other important proteins like METTL3 and METTL14. WTAP does not bind RNA directly but helps its partners achieve target specificity during gene expression processes. The presence of WTAP within the complex stabilizes and coordinates the activity of the entire methyltransferase assembly.

Pathways

The m6A methylation WTAP facilitates affects various pathways including mRNA splicing and degradation. This process intricately connects WTAP to the regulation of the JAK-STAT signaling pathway where its activity influences inflammatory responses and cell proliferation. Additionally WTAP has ties to the Wnt/β-catenin pathway collaborating with proteins like METTL3 to regulate gene expression involved in cell fate determination and embryogenesis.

Associated diseases and disorders

Research shows WTAP has connections to various cancers such as hepatocellular carcinoma due to its role in mRNA processing and stability. This association often involves its interaction with other oncogenic proteins like METTL3 leading to aberrant gene expression patterns. WTAP also has relevance in acute myeloid leukemia where it influences mRNA maturation and impacts cell differentiation and proliferation.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

7 product images

  • Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351), expandable thumbnail

    Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351)

    This data was developed using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

    In Western blot, anti-WTAP antibody (Anti-WTAP antibody [EPR18744] ab195380) as pan control staining at 1/1000 dilution.

    In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

    All lanes: Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] (Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350) at 1/1000 dilution

    Lane 1: Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate (untreated membrane) at 20 µg

    Lane 2: Wild-type HAP1 whole cell lysate (Alkaline phosphatase treated membrane) 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

    Developed using the ECL technique.

    Observed band size: 55 kDa

    Exposure time: 180s

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

    In Western blot, anti-WTAP antibody (Anti-WTAP antibody [EPR18744] ab195380) as pan control staining at 1/1000 dilution.

    In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

  • Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351), expandable thumbnail

    Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351)

    This data was developed using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression of WTAP (phospho S341) is upregulated in response to PMA treatment (PMID:33217317).

    In Western blot, anti-WTAP antibody (Anti-WTAP antibody [EPR18744] ab195380) as pan control staining at 1/1000 dilution.

    In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

    All lanes: Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] (Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350) at 1/1000 dilution

    Lane 1: Untreated Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 20 µg

    Lane 2: HAP1 treated with /ml PMA for 30 minutes whole cell lysate at 20 µg

    Lane 3: Untreated WTAP knockout HAP1 whole cell lysate at 20 µg

    Lane 4: WTAP knockout HAP1 treated with /ml PMA for 30 minutes 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

    Observed band size: 55 kDa

    Exposure time: 180s

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    The expression of WTAP (phospho S341) is upregulated in response to PMA treatment (PMID:33217317).

    In Western blot, anti-WTAP antibody (Anti-WTAP antibody [EPR18744] ab195380) as pan control staining at 1/1000 dilution.

    In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
    Exposure time: 180 seconds

  • Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351), expandable thumbnail

    Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351)

    This data was developed using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350, the same antibody clone in a different buffer formulation.
    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    The samples were run on a Bis-Tris gel under reducing conditions.

    Western blot: Anti-WTAP (phospho S341)antibody (Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.

    The identity of the lower MW at approximately 35KD is unknown.

    To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescentwestern blot (TBS-based) blocking solution before incubation with primaryantibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

    All lanes: Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] (Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350) at 1/1000 dilution

    All lanes: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rat IgG H&L (Goat Anti-Rat IgG H&L ab97053) at 1/100000 dilution

    Observed band size: 55 kDa

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    The samples were run on a Bis-Tris gel under reducing conditions.

    Western blot: Anti-WTAP (phospho S341)antibody (Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.

    The identity of the lower MW at approximately 35KD is unknown.

    To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescentwestern blot (TBS-based) blocking solution before incubation with primaryantibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351)

    This data was developed using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350, the same antibody clone in a different buffer formulation.
    Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling WTAP (phospho S341) with Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 at 1/500 (1.044 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human endometrium without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin.

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351)

    This data was developed using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350, the same antibody clone in a different buffer formulation.
    Immunohistochemical analysis of paraffin-embedded Human endometrial ca tissue labeling WTAP (phospho S341) with Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 at 1/500 (1.044 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human endometrial carcinoma without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B).The section was incubated with Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunoprecipitation - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351), expandable thumbnail

    Immunoprecipitation - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351)

    This data was developed using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350, the same antibody clone in a different buffer formulation.
    WTAP (phospho S341) was immunoprecipitated from 0.35 mg Parental HAP1 (wildtype control human chronic myelogenous leukemia near-haploid cell line) whole cell lysate with Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: Parental HAP1 (wildtype control human chronic myelogenous leukemia near-haploid cell line) whole cell lysate 4 ug

    Lane 2: abab309350 IP in Parental HAP1 (wildtype control human chronic myelogenous leukemia near-haploid cell line) whole cell lysate

    Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 in parental HAP1whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 180 seconds

    All lanes: Immunoprecipitation - Anti-WTAP (phospho S341) antibody [EPR27039-57] (Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350) at 1/30 dilution

    All lanes: Parental HAP1 whole cell lysate at 4 µg

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Observed band size: 55 kDa

    Exposure time: 180s

    WTAP (phospho S341) was immunoprecipitated from 0.35 mg Parental HAP1 (wildtype control human chronic myelogenous leukemia near-haploid cell line) whole cell lysate with Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: Parental HAP1 (wildtype control human chronic myelogenous leukemia near-haploid cell line) whole cell lysate 4 ug

    Lane 2: Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 IP in Parental HAP1 (wildtype control human chronic myelogenous leukemia near-haploid cell line) whole cell lysate

    Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 in parental HAP1whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 180 seconds

  • Dot Blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351), expandable thumbnail

    Dot Blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] - BSA and Azide free (ab309351)

    This data was developed using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350, the same antibody clone in a different buffer formulation.

    Dot blot analysis of WTAP (phospho S341) using Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350 at 1:1000 (0.522 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

    Exposure time: 180 seconds
    Blocking and diluting buffer and concentration: 5% NFDM/TBST

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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