重组Anti-HNF-4-alpha抗体[EPR16885] - ChIP Grade (ab181604)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16885] to HNF-4-alpha - ChIP Grade
- Suitable for: ChIC/CUT&RUN-seq, ChIP, IHC-P, WB, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-HNF-4-alpha抗体[EPR16885] - ChIP Grade
参阅全部 HNF-4-alpha 一抗 -
描述
兔单克隆抗体[EPR16885] to HNF-4-alpha - ChIP Grade -
宿主
Rabbit -
经测试应用
适用于: ChIC/CUT&RUN-seq, ChIP, IHC-P, WB, IPmore details -
种属反应性
与反应: Mouse, Rat, 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: HepG2 and SW480 whole cell lysates; Human fetal liver, colon and fetal kidney lysates; mouse and rat liver lysates. IHC-P: Human liver, Human colon, mouse liver and rat colon tissues. IP: HepG2 whole cell extract. ChIP: HepG2 whole cell extract. ChIC/CUT&RUN-Seq: HepG2 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
Preservative: 0.01% Sodium azide
Constituents: 59% 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
EPR16885 -
同种型
IgG -
研究领域
- Epigenetics and Nuclear Signaling
- Transcription
- Polymerase associated factors
- Pol II Transcription
- Other
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipases
相关产品
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab181604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 |
---|---|---|
ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5 µg |
|
ChIP | (1) |
Use 2 µg for 25 µg of chromatin.
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IHC-P | (1) |
1/2000. 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 53 kDa (predicted molecular weight: 53 kDa).
|
|
IP |
1/70.
|
说明 |
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ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5 µg |
ChIP
Use 2 µg for 25 µg of chromatin. |
IHC-P
1/2000. 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 53 kDa (predicted molecular weight: 53 kDa). |
IP
1/70. |
靶标
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功能
Transcriptionally controlled transcription factor. Binds to DNA sites required for the transcription of alpha 1-antitrypsin, apolipoprotein CIII, transthyretin genes and HNF1-alpha. May be essential for development of the liver, kidney and intestine. -
疾病相关
Defects in HNF4A are the cause of maturity-onset diabetes of the young type 1 (MODY1) [MIM:125850]; also symbolized MODY-1. MODY is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease. -
序列相似性
Belongs to the nuclear hormone receptor family. NR2 subfamily.
Contains 1 nuclear receptor DNA-binding domain. -
翻译后修饰
Phosphorylated on tyrosine residue(s); phosphorylation is important for its DNA-binding activity. Phosphorylation may directly or indirectly play a regulatory role in the subnuclear distribution. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 3172 Human
- Entrez Gene: 15378 Mouse
- Entrez Gene: 25735 Rat
- Omim: 600281 Human
- SwissProt: P41235 Human
- SwissProt: P49698 Mouse
- SwissProt: P22449 Rat
- Unigene: 116462 Human
see all -
别名
- FLJ39654 antibody
- FRTS4 antibody
- Hepatic nuclear factor 4 alpha antibody
see all
图片
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ChIC/CUT&RUN sequencing – Recombinant Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (Human liver hepatocellular carcinoma cell line) cells and 5 µg of ab181604 [EPR16885]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab181604 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
ABCC6_NC1 is negative control
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All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/10000 dilution
Lane 1 : HepG2 whole cell lysate
Lane 2 : HEK293 whole cell lysate
Lane 3 : Mouse liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Predicted band size: 53 kDaThis blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour before being incubated with ab181604 (rabbit monoclonal [EPR16885] to HNF-4-alpha; dilution 1:10000) and loading control ab18058 (mouse monoclonal [SPM227] to Vinculin; dilution 1:10000) at 4°C overnight. Antibody binding was detected with ab216773 (Goat anti-Rabbit IgG H&L (IRDye® 800CW); green; dilution 1:20000) and ab216776 (Goat anti-Mouse IgG H&L (IRDye® 680RD), red; dilution 1:20000) for 1 hour at room temperature before imaging.
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Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution + Full length mouse HNF-4-gamma recombinant protein at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/10000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution + SW480 (Human colon adenocarcinoma cell line) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human colon lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
Lane 1 : Mouse liver lysate
Lane 2 : Rat liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Exposure time: 15 secondsThis antibody can recognize 2 isoforms in mouse and rat. The predicted MW are 53KDa and 52 KDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human liver 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.
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Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of Human colon 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.
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Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on mouse liver 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.
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Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of rat colon 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.
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HNF-4 alpha was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell extract with ab181604 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab181604 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HepG2 whole cell extract 10 µg (Input).
Lane 2: ab181604 IP in HepG2 whole cell extract.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181604 in HepG2 whole cell extract.
Blocking and dilution buffer and concentration: 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 (43)
ab181604 被引用在 43 文献中.
- Zhang J et al. Naringenin in Si-Ni-San formula inhibits chronic psychological stress-induced breast cancer growth and metastasis by modulating estrogen metabolism through FXR/EST pathway. J Adv Res 47:189-207 (2023). PubMed: 35718080
- Park H et al. Discovery of 3,7-dimethoxyflavone that inhibits liver fibrosis based on dual mechanisms of antioxidant and inhibitor of activated hepatic stellate cell. Free Radic Biol Med 204:195-206 (2023). PubMed: 37146699
- Luo H et al. The HNF4A-CHPF pathway promotes proliferation and invasion through interactions with MAD1L1 in glioma. Aging (Albany NY) 15:11052-11066 (2023). PubMed: 37851364
- Lei X et al. Epithelial HNF4A shapes the intraepithelial lymphocyte compartment via direct regulation of immune signaling molecules. J Exp Med 219:N/A (2022). PubMed: 35792863
- Li Y et al. Macrophages activated by hepatitis B virus have distinct metabolic profiles and suppress the virus via IL-1β to downregulate PPARα and FOXO3. Cell Rep 38:110284 (2022). PubMed: 35081341