Rabbit Recombinant Monoclonal WTAP antibody. Suitable for IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 47 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | WB | IHC-P | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Primates | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/40 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Primates | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes Milk recommended as blocking agent. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Milk recommended as blocking agent. |
Species Primates | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Primates | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Primates | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/600 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Primates | Dilution info - | Notes - |
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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).
KIAA0105, KIAA0105, WTAP, Pre-mRNA-splicing regulator WTAP, Female-lethal(2)D homolog, WT1-associated protein, Wilms tumor 1-associating protein, hFL(2)D
Rabbit Recombinant Monoclonal WTAP antibody. Suitable for IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 47 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR18744
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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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.
Lane 1: Wild-type HAP1 whole cell lysate (20 μg)
Lane 2: WTAP knockout HAP1 whole cell lysate (20 μg)
Lane 3: MOLT-4 whole cell lysate (20 μg)
Lane 4: K562 whole cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab195380 observed at 50 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab195380 was shown to specifically react with WTAP in wild-type HAP1 cells as signal was lost in WTAP knockout cells. Wild-type and WTAP knockout samples were subjected to SDS-PAGE. ab195380 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-WTAP antibody [EPR18744] (ab195380)
Predicted band size: 44 kDa
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling WTAP with ab195380 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Jurkat cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab195380 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Blocking/Dilution buffer: 5% NFDM/TBST.
The observed molecular weight is consistent with what has been described in the literature (PMID: 17088532).
All lanes: Western blot - Anti-WTAP antibody [EPR18744] (ab195380) at 1/1000 dilution
Lane 1: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 2: K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate at 20 µg
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
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.
Predicted band size: 44 kDa
Observed band size: 50 kDa
Exposure time: 3min
Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling WTAP with ab195380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on human endometrium is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling WTAP with ab195380 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
The observed molecular weight is consistent with what has been described in the literature (PMID: 17088532).
All lanes: Western blot - Anti-WTAP antibody [EPR18744] (ab195380) at 1/1000 dilution
Lane 1: Human kidney lysate at 10 µg
Lane 2: Human thymus lysate at 10 µg
Lane 3: Human lung lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 44 kDa
Observed band size: 50 kDa
Exposure time: 3min
WTAP was immunoprecipitated from 0.35 mg of Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate with ab195380 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab195380 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: Jurkat whole cell lysate 10μg (Input).
Lane 2: ab195380 IP in Jurkat whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab195380 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
All lanes: Immunoprecipitation - Anti-WTAP antibody [EPR18744] (ab195380)
Predicted band size: 44 kDa
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling WTAP with ab195380 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab195380 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling WTAP with ab195380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on human liver cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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 (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
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
The expression of WTAP (phospho S341) is upregulated in response to PMA treatment (PMID:33217317).
In Western blot, anti-WTAP antibody (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
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
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
In Western blot, Anti-WTAP antibody [EPR18744] - Loading Control (ab195380) staining at 1/1000 dilution.
All lanes: Western blot - Anti-WTAP (phospho S306) antibody [EPR27038-188] (Anti-WTAP (phospho S306) antibody [EPR27038-188] ab316275) at 1/1000 dilution
Lane 1: A375 (human malignant melanoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg
Lane 2: A375 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 55 kDa
Exposure time: 26s
In Western blot, Anti-WTAP antibody [EPR18744] - Loading Control (ab195380) staining at 1/1000 dilution.
All lanes: Western blot - Anti-WTAP (phospho S306) antibody [EPR27038-188] (Anti-WTAP (phospho S306) antibody [EPR27038-188] ab316275) at 1/1000 dilution
Lane 1: Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate (untreated membrane) at 20 µg
Lane 2: Jurkat whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 55 kDa
Exposure time: 26s
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