重组Anti-KLF4抗体[EPR20753-25] (ab214666)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20753-25] to KLF4
- Suitable for: IHC-P, WB, ICC/IF, IP, IHC - Wholemount
- Reacts with: Mouse, Rat
Related conjugates and formulations
概述
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产品名称
Anti-KLF4抗体[EPR20753-25]
参阅全部 KLF4 一抗 -
描述
兔单克隆抗体[EPR20753-25] to KLF4 -
宿主
Rabbit -
经测试应用
适用于: IHC-P, WB, ICC/IF, IP, IHC - Wholemountmore details -
种属反应性
与反应: Mouse, Rat -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: F9 and ES-D3 whole cell lysate; Mouse and rat skin and mouse colon tissue lysate. IHC-P: Mouse and rat colon tissue. ICC/IF: F9 cells. IP: F9 whole cell lysate.
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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, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR20753-25 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab214666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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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 54 kDa.
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ICC/IF |
1/1000.
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IP |
1/30.
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IHC - Wholemount | (1) |
Use at an assay dependent concentration.
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说明 |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 54 kDa. |
ICC/IF
1/1000. |
IP
1/30. |
IHC - Wholemount
Use at an assay dependent concentration. |
靶标
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功能
Transcription factor; can act both as activator and as repressor. Binds the 5'-CACCC-3' core sequence. Binds to the promoter region of its own gene and can activate its own transcription. Regulates the expression of key transcription factors during embryonic development. Plays an important role in maintaining embryonic stem cells, and in preventing their differentiation. Required for establishing the barrier function of the skin and for postnatal maturation and maintenance of the ocular surface. Involved in the differentiation of epithelial cells and may also function in skeletal and kidney development. Contributes to the down-regulation of p53/TP53 transcription. -
序列相似性
Belongs to the krueppel C2H2-type zinc-finger protein family.
Contains 3 C2H2-type zinc fingers. -
结构域
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 16600 Mouse
- Entrez Gene: 114505 Rat
- SwissProt: Q60793 Mouse
- Unigene: 4325 Mouse
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别名
- Endothelial Kruppel like zinc finger protein antibody
- Epithelial zinc finger protein EZF antibody
- EZF antibody
see all
图片
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Several embryos at the mid-blastocyst stage (~E3.75) were tested. The embryos were fixed for 10 mins at room temperature in 4% PFA. Fixed embryos were washed for 5 mins in PBS + 0.1% Triton X-100 (PBX) at room temperature. The embryos were then permeabilized in PBS + 0.5% Triton X-100 + 100mM Glycine for 5 mins at room temperature and washed again in PBX for 5 mins at room temperature. Blocking was carried out for 1 hour at room temperature in PBS + 2% horse serum and then the embryos were incubated overnight in ab214666 (1/1000) diluted in blocking solution. The next day embryos were washed three times for 5 mins each in PBX, before blocking again for 1 hour at room temperature. Then incubated in Donkey anti rabbit Alexa Fluor® 488 secondary antibodies (1/500) diluted in blocking solution for 1 hour at 4ºC. This was followed by 2 washes of 5 mins each in PBX. The embryos were then incubated in PBS + 5μg/ml DAPI for 10 min, or until mounting for imaging. Images were acquired on a laser-scanning confocal microscope system. Displayed image shows maximum intensity projection of 5 optical sections acquired at 1μm increments. Scale bar: 20 μm
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Lanes 1-4 : Anti-KLF4 antibody [EPR20753-25] (ab214666) at 1/1000 dilution
Lane 5 : Anti-KLF4 antibody [EPR20753-25] (ab214666) at 1/5000 dilution
Lane 1 : F9 (mouse embryonal carcinoma epithelial cell), whole cell lysate
Lane 2 : Mouse skin tissue lysate
Lane 3 : Mouse colon tissue lysate
Lane 4 : Rat skin tissue lysate
Lane 5 : ES D3 (mouse embryonic stem cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Lane 5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?Exposure times:
Lanes 1-2: 3 minutes
Lane 3-4: 92 seconds
Lane 5: 48 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
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KLF4 was immunoprecipitated from 0.35 mg F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysate using ab214666 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab214666 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary at 1/5000 dilution.
Lane 1: F9 whole cell lysate 10 µg (input).
Lane 2: ab214666 IP in F9 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214666 in F9 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
Fresh lysate was required and IP test was conducted immediately. Incubation time was shortened from overnight to 2h.
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Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling KLF4 with ab214666 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Nuclear staining on epithelial cells of rat colon (PMID: 21070761, PMID: 19109406) is observed. Counterstained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling KLF4 with ab214666 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Nuclear staining on epithelial cells of mouse colon (PMID: 21070761, PMID: 19109406) is observed. Counterstained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labeling KLF4 with ab214666 at 1/100 dilution, followed by a AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green). Confocal image showing nuclear staining in F9 cell line is observed. The nuclear counterstain was DAPI (Blue). Tubulin was counterstained using an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889, Red) at 1.200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000.
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (14)
ab214666 被引用在 14 文献中.
- Huang N et al. Plasma exosomes confer hypoxic pulmonary hypertension by transferring LOX-1 cargo to trigger phenotypic switching of pulmonary artery smooth muscle cells. Biochem Pharmacol 207:115350 (2023). WB ; Rat . PubMed: 36435201
- Eenjes E et al. Functional Characterization and Visualization of Esophageal Fibroblasts Using Organoid Co-Cultures. J Vis Exp N/A:N/A (2023). IHC - Wholemount ; Mouse . PubMed: 36688545
- Huang Z et al. microRNA-130b May Induce Cerebral Vasospasm after Subarachnoid Hemorrhage via Modulating Kruppel-like Factor 4. Mol Cell Biol N/A:1-16 (2023). WB, IHC-P ; Rat . PubMed: 37381993
- Wang X et al. The KLF4-STAT5 axis promotes pancreatic fibrosis in mice with caerulein-induced chronic pancreatitis. Exp Anim N/A:N/A (2023). WB, IHC-P, ChIP ; Mouse . PubMed: 36948613
- Wang J et al. KLF4 Alleviates Hypertrophic Scar Fibrosis by Directly Activating BMP4 Transcription. Int J Biol Sci 18:3324-3336 (2022). PubMed: 35637963