重组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
概述
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产品名称
Anti-GATA3抗体[EPR16651] - ChIP Grade
参阅全部 GATA3 一抗 -
描述
兔单克隆抗体[EPR16651] to GATA3 - ChIP Grade -
宿主
Rabbit -
特异性
IHC-P is suitable in Human, and may not be suitable for use in Mouse samples.
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经测试应用
适用于: Flow Cyt (Intra), ChIP, WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- 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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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
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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, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR16651 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-GATA3 antibody [EPR16651] (ab208895)
- Alexa Fluor® 647 Anti-GATA3 antibody [EPR16651] (ab208896)
- Alexa Fluor® 555 Anti-GATA3 antibody [EPR16651] (ab210672)
- Anti-GATA3 antibody [EPR16651] - BSA and Azide free (ab214804)
- APC Anti-GATA3 antibody [EPR16651] (ab225418)
- PE Anti-GATA3 antibody [EPR16651] (ab225419)
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab199428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 |
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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 | (7) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
ICC/IF | (2) |
1/250.
|
说明 |
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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. |
靶标
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功能
Transcriptional activator which binds to the enhancer of the T-cell receptor alpha and delta genes. Binds to the consensus sequence 5'-AGATAG-3'. -
组织特异性
T-cells and endothelial cells. -
疾病相关
Defects in GATA3 are the cause of hypoparathyroidism with sensorineural deafness and renal dysplasia (HDR) [MIM:146255]; also known as Barakat syndrome. -
序列相似性
Contains 2 GATA-type zinc fingers. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- 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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形式
There are 2 isoforms produced by alternative splicing. -
别名
- GATA 3 antibody
- GATA binding factor 3 antibody
- GATA binding protein 3 antibody
see all
图片
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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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GATA3 antibody [EPR16651] - ChIP Grade (ab199428)
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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GATA3 antibody [EPR16651] - ChIP Grade (ab199428)
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.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (46)
ab199428 被引用在 46 文献中.
- 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