Anti-WSTF 抗体 [EP1704Y] (ab51256)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1704Y] to WSTF
- Suitable for: IHC-P, WB, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-WSTF antibody [EP1704Y]
WSTF 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1704Y] to WSTF -
由来種
Rabbit -
アプリケーション
適用あり: IHC-P, WB, Flow Cyt (Intra), ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human WSTF (C terminal). The exact sequence is proprietary.
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ポジティブ・コントロール
- WB: Wild-type HAP1, HeLa (ab150035), and PC12 cell lysates. Mouse and Rat testis. B16-F0, MCF7, HeLa whole cell lysates. Flow cyt: HeLa cells; IHC: rat cardiac muscle tissue, mouse cardiac muscle tissue, human breast carcinoma; ICC/IF: HeLa cells.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1704Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab51256の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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IHC-P |
1/700. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB | (2) |
1/15000 - 1/20000. Detects a band of approximately 185 kDa (predicted molecular weight: 171 kDa).
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Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/50.
For unpurified format use at 1/100 to 1/250 dilution |
特記事項 |
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IHC-P
1/700. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
1/15000 - 1/20000. Detects a band of approximately 185 kDa (predicted molecular weight: 171 kDa). |
Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/50. For unpurified format use at 1/100 to 1/250 dilution |
ターゲット情報
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機能
Atypical tyrosine-protein kinase that plays a central role in chromatin remodeling and acts as a transcription regulator. Involved in DNA damage response by phosphorylating 'Tyr-142' of histone H2AX (H2AXY142ph). H2AXY142ph plays a central role in DNA repair and acts as a mark that distinguishes between apoptotic and repair responses to genotoxic stress. Essential component of the WICH complex, a chromatin remodeling complex that mobilizes nucleosomes and reconfigures irregular chromatin to a regular nucleosomal array structure. The WICH complex regulates the transcription of various genes, has a role in RNA polymerase I and RNA polymerase III transcription, mediates the histone H2AX phosphorylation at 'Tyr-142', and is involved in the maintenance of chromatin structures during DNA replication processes. In the complex, it mediates the recruitment of the WICH complex to replication foci during DNA replication. Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene. In the WINAC complex, plays an essential role by targeting the complex to acetylated histones, an essential step for VDR-promoter association. -
組織特異性
Ubiquitously expressed with high levels of expression in heart, brain, placenta, skeletal muscle and ovary. -
関連疾患
Note=BAZ1B is located in the Williams-Beuren syndrome (WBS) critical region. WBS results from a hemizygous deletion of several genes on chromosome 7q11.23, thought to arise as a consequence of unequal crossing over between highly homologous low-copy repeat sequences flanking the deleted region. Haploinsufficiency of BAZ1B may be the cause of certain cardiovascular and musculo-skeletal abnormalities observed in the disease. -
配列類似性
Belongs to the WAL family. BAZ1B subfamily.
Contains 1 bromo domain.
Contains 1 DDT domain.
Contains 1 PHD-type zinc finger.
Contains 1 WAC domain. -
発生段階
Expressed at equal levels in 19-23 weeks old fetal tissues. -
ドメイン
The N-terminal part (1-345), including the WAC domain and the C motif, mediates the tyrosine-protein kinase activity.
The bromo domain mediates the specific interaction with acetylated histones. -
翻訳後修飾
Phosphorylated upon DNA damage, probably by ATM or ATR. -
細胞内局在
Nucleus. Accumulates in pericentromeric heterochromatin during replication. Targeted to replication foci throughout S phase via its association with PCNA. - Information by UniProt
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参照データベース
- Entrez Gene: 9031 Human
- Entrez Gene: 22385 Mouse
- Entrez Gene: 368002 Rat
- Omim: 605681 Human
- SwissProt: Q9UIG0 Human
- SwissProt: Q9Z277 Mouse
- Unigene: 647016 Human
- Unigene: 40331 Mouse
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別名
- baz1b antibody
- BAZ1B_HUMAN antibody
- Bromodomain adjacent to zinc finger domain protein 1B antibody
see all
画像
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Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling WSTF with Purified ab51256 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling WSTF with Purified ab51256 at 1:50 dilution (2.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cardiac muscle tissue sections labeling WSTF with Purified ab51256 at 1:700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue sections labeling WSTF with Purified ab51256 at 1:700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling WSTF with Purified ab51256 at 1:700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
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All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/5000 dilution
Lane 1 : B16-F0 (Mouse melanoma epithelial cell-like) whole cell lysate
Lane 2 : Mouse testis lysate
Lane 3 : Rat testis lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 171 kDa
Observed band size: 185 kDa why is the actual band size different from the predicted?Exposure time
Lane 1: 10 seconds
Lane 2: 40 seconds
Lane 3: 180 seconds
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All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/15000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BAZ1B knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 171 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab51256 observed at 175 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab51256 was shown to react with WSTF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264907 (knockout cell lysate ab257370) was used. Wild-type HeLa and WSTF knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51256 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 15000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/20000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 171 kDaExposure time:
Left image: 180 seconds
Right image: 40 seconds
We recommend to use ab109439 for WSTF Western Blot testing because ab51256 detects nonspecific bands.
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All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/15000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : WSTF knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : PC12 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 171 kDaLanes 1 - 4: Merged signal (red and green). Green - ab51256 observed at 175 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab51265 was shown to recognize WSTF in wilt-type cells along with additional cross-reactive bands as signal was lost in WSTF knockout samples. Wild-type and WSTF knockout samples were subjected to SDS-PAGE. ab51256 and ab18058 (loading control to vinculin) were diluted 1/15 000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (8)
ab51256 は 8 報の論文で使用されています。
- D'Alesio C et al. RNAi screens identify CHD4 as an essential gene in breast cancer growth. Oncotarget 7:80901-80915 (2016). WB . PubMed: 27779108
- Sun H et al. BAZ1B in Nucleus Accumbens Regulates Reward-Related Behaviors in Response to Distinct Emotional Stimuli. J Neurosci 36:3954-61 (2016). PubMed: 27053203
- Liu Y et al. KRASG12 mutant induces the release of the WSTF/NRG3 complex, and contributes to an oncogenic paracrine signaling pathway. Oncotarget 7:53153-53164 (2016). PubMed: 27449290
- Ashton NW et al. Novel insight into the composition of human single-stranded DNA-binding protein 1 (hSSB1)-containing protein complexes. BMC Mol Biol 17:24 (2016). WB . PubMed: 27938330
- Sun H et al. ACF chromatin-remodeling complex mediates stress-induced depressive-like behavior. Nat Med 21:1146-53 (2015). WB . PubMed: 26390241
- Vidi PA et al. NuMA promotes homologous recombination repair by regulating the accumulation of the ISWI ATPase SNF2h at DNA breaks. Nucleic Acids Res 42:6365-79 (2014). ICC/IF ; Human . PubMed: 24753406
- Sánchez-Molina S et al. Role for hACF1 in the G2/M damage checkpoint. Nucleic Acids Res 39:8445-56 (2011). WB . PubMed: 21745822
- Yoshimura K et al. Distinct function of 2 chromatin remodeling complexes that share a common subunit, Williams syndrome transcription factor (WSTF). Proc Natl Acad Sci U S A 106:9280-5 (2009). ICC/IF ; Mouse . PubMed: 19470456