Anti-GATA3 抗体 [EPR16651] - ChIP Grade (ab199428)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16651] to GATA3 - ChIP Grade
- Suitable for: Flow Cyt (Intra), ChIP, WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-GATA3 antibody [EPR16651] - ChIP Grade
GATA3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR16651] to GATA3 - ChIP Grade -
由来種
Rabbit -
特異性
IHC-P is suitable in Human, and may not be suitable for use in Mouse samples.
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アプリケーション
適用あり: Flow Cyt (Intra), ChIP, WB, IHC-P, ICC/IFmore details -
種交差性
交差種: Mouse, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: RAW 264.7 whole cell lysate (ab7187), SH-SY5Y cell extracts. EL4, HAP1 and Jurkat whole cell lysates. Mouse brain lysate. IHC-P: Human neuroblastoma and Human breast carcinoma tissue. ICC/IF: SH-SY5Y cells. Flow Cyt (intra): Jurkat cells. ChIP: Chromatin prepared from MCF7 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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 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
EPR16651 -
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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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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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ab199428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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Flow Cyt (Intra) |
1/500.
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ChIP |
Use 2 µg for 25 µg of chromatin.
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WB |
1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
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IHC-P | (8) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (2) |
1/250.
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特記事項 |
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Flow Cyt (Intra)
1/500. |
ChIP
Use 2 µg for 25 µg of chromatin. |
WB
1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/250. |
ターゲット情報
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機能
Transcriptional activator which binds to the enhancer of the T-cell receptor alpha and delta genes. Binds to the consensus sequence 5'-AGATAG-3'. -
組織特異性
T-cells and endothelial cells. -
関連疾患
Defects in GATA3 are the cause of hypoparathyroidism with sensorineural deafness and renal dysplasia (HDR) [MIM:146255]; also known as Barakat syndrome. -
配列類似性
Contains 2 GATA-type zinc fingers. -
細胞内局在
Nucleus. - Information by UniProt
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参照データベース
- Entrez Gene: 2625 Human
- Entrez Gene: 14462 Mouse
- Omim: 131320 Human
- SwissProt: P23771 Human
- SwissProt: P23772 Mouse
- Unigene: 524134 Human
- Unigene: 313866 Mouse
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製品の状態
There are 2 isoforms produced by alternative splicing. -
別名
- GATA 3 antibody
- GATA binding factor 3 antibody
- GATA binding protein 3 antibody
see all
画像
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All lanes : Anti-GATA3 antibody [EPR16651] - ChIP Grade (ab199428) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : EL4 (mouse lymphoma T lymphocyte) 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 48 kDa
Observed band size: 48 kDaExposure
Lane1: 26 seconds
Lane 2: 15 seconds
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Chromatin was prepared from MCF7 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25μg of chromatin, 2μg of ab199428 (blue), and 20μl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the IgG control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach). Primers and probes are located in the first kb of the transcribed region.
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All lanes : Anti-GATA3 antibody [EPR16651] - ChIP Grade (ab199428) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GATA3 knockout HAP1 whole cell lysate
Lane 3 : Jurkat whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 48 kDaLanes 1 - 3: Merged signal (red and green). Green - ab199428 observed at 48 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab199428 was shown to recognize GATA3 in wild-type HAP1 cells as signal was lost at the expected MW in GATA3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GATA3 knockout samples were subjected to SDS-PAGE. Ab199428 and ab9484 (Mouse anti-GAPDH 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 1/20000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling GATA3 with ab199428 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human breast carcinoma tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y cells (Human neuroblastoma from bone marrow cells) labeling GATA3 with ab199428 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on SH-SY5Y cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab199428 at 1/250 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
Intracellular Flow Cytometry analysis of Jurkat cells labelling GATA3 with ab199428 at 1/500 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluorr® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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Immunohistochemical analysis of paraffin-embedded Human neuroblastoma tissue labeling GATA3 with ab199428 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human neuroblastoma tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Anti-GATA3 antibody [EPR16651] - ChIP Grade (ab199428) at 1/1000 dilution + SH-SY5Y (Human neuroblastoma from bone marrow cells) cell extract at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 48 kDa
Observed band size: 48 kDa
Exposure time: 1 minuteBlocking/dilution buffer: 5% NFDM/TBST.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (46)
ab199428 は 46 報の論文で使用されています。
- Moya-Jódar M et al. Revealing cell populations catching the early stages of human embryo development in naive pluripotent stem cell cultures. Stem Cell Reports 18:64-80 (2023). PubMed: 36563688
- Zhang Y et al. E239K mutation abolishes the suppressive effects of lysine-specific demethylase 1 on migration and invasion of MCF7 cells. Cancer Sci 113:489-499 (2022). PubMed: 34839571
- Chou WC et al. A common variant in 11q23.3 associated with hyperlipidemia is mediated by the binding and regulation of GATA4. NPJ Genom Med 7:4 (2022). PubMed: 35046404
- Yu L & Li J Punicalagin Alleviates Aged Bronchial Asthma by Inhibiting Th2 Differentiation through IL-4/STAT6 and Jagged1/Notch Pathways. J Healthc Eng 2022:1184677 (2022). PubMed: 35140898
- Chen P et al. Gata3 Silencing Is Involved in Neuronal Differentiation and Its Abnormal Expression Impedes Neural Activity in Adult Retinal Neurocytes. Int J Mol Sci 23:N/A (2022). PubMed: 35269648