重组Anti-WTAP抗体[EPR18744] (ab195380)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18744] to WTAP
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
-
产品名称
Anti-WTAP抗体[EPR18744]
参阅全部 WTAP 一抗 -
描述
兔单克隆抗体[EPR18744] to WTAP -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details -
种属反应性
与反应: Human
预测可用于: Pig, Non human primates不与反应: Mouse -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- WB: Jurkat, K562, HeLa and HepG2 whole cell lysates; human kidney, thymus and lung lysates. IHC-P: Human endometrium and liver cancer tissues. ICC/IF: Jurkat and HeLa cells. Flow Cyt (intra): Jurkat cells. IP: Jurkat whole cell lysate.
-
常规说明
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.
性能
-
形式
Liquid -
存放说明
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. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18744 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab195380于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
Flow Cyt (Intra) |
1/600.
|
|
ICC/IF |
1/500.
|
|
IP |
1/40.
|
|
IHC-P | (3) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
WB | (2) |
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 44 kDa).
Milk recommended as blocking agent. |
说明 |
---|
Flow Cyt (Intra)
1/600. |
ICC/IF
1/500. |
IP
1/40. |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 44 kDa). Milk recommended as blocking agent. |
靶标
-
功能
Regulates G2/M cell-cycle transition by binding to the 3' UTR of CCNA2, which enhances its stability. Impairs WT1 DNA-binding ability and inhibits expression of WT1 target genes. May be involved in mRNA splicing regulation. -
组织特异性
Ubiquitously expressed. -
序列相似性
Belongs to the fl(2)d family. -
翻译后修饰
Phosphorylated upon DNA damage, probably by ATM or ATR. -
细胞定位
Nucleus > nucleolus. - Information by UniProt
-
数据库链接
- Entrez Gene: 9589 Human
- Entrez Gene: 100157533 Pig
- Omim: 605442 Human
- SwissProt: Q15007 Human
- Unigene: 446091 Human
-
别名
- DKFZp686F20131 antibody
- Female-lethal(2)D homolog antibody
- FL2D_HUMAN antibody
see all
图片
-
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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
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) (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 (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (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) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
All lanes : Anti-WTAP antibody [EPR18744] (ab195380) at 1/1000 dilution
Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) 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
Developed using the ECL technique.
Predicted band size: 44 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.The observed molecular weight is consistent with what has been described in the literature (PMID: 17088532).
-
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) (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) (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 (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.
-
All lanes : Anti-WTAP antibody [EPR18744] (ab195380) at 1/1000 dilution
Lane 1 : Human kidney lysate
Lane 2 : Human thymus lysate
Lane 3 : Human lung lysate
Lysates/proteins at 10 µg per lane.
Secondary
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 why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The observed molecular weight is consistent with what has been described in the literature (PMID: 17088532).
-
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) (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 (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (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) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (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) (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) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
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) (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 (ab172730) instead of ab195380 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
实验方案
数据表及文件
-
SDS download
-
Datasheet download
Certificate of Compliance
文献 (38)
ab195380 被引用在 38 文献中.
- Polenkowski M et al. THOC5 complexes with DDX5, DDX17, and CDK12 to regulate R loop structures and transcription elongation rate. iScience 26:105784 (2023). PubMed: 36590164
- Chen Y et al. Qinzhu Liangxue inhibits IL-6-induced hyperproliferation and inflammation in HaCaT cells by regulating METTL14/SOCS3/STAT3 axis. J Ethnopharmacol 317:116809 (2023). WB ; Human . PubMed: 37336334
- Tao CJ et al. High expression of WTAP is related to poor prognosis in nasopharyngeal carcinoma. Neoplasma 70:229-239 (2023). IHC-P ; Human . PubMed: 36964720
- Huang L et al. WTAP regulates autophagy in colon cancer cells by inhibiting FLNA through N6-methyladenosine. Cell Adh Migr 17:1-13 (2023). WB ; Human . PubMed: 36849408
- Lin Z et al. N6-methyladenosine (m6A) methyltransferase WTAP-mediated miR-92b-5p accelerates osteoarthritis progression. Cell Commun Signal 21:199 (2023). WB ; Human . PubMed: 37563688