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主营:分子类,蛋白类,抗体类,生化类试剂
℡ 4000-520-616
℡ 4000-520-616
RayBiotech/Human HNF-1-alpha Transcription Factor Activity Assay/1 Plate Kit/TFEH-HNF1a-1
产品编号:TFEH-HNF1a-1
市  场 价:¥0.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:待定
品      牌: RayBiotech
公      司:RayBiotech
公司分类:
RayBiotech/Human HNF-1-alpha Transcription Factor Activity Assay/1 Plate Kit/TFEH-HNF1a-1
商品介绍

Antigen Information

Accession Number:
  • P20823
Gene ID:
  • 6927
Gene Symbols:
  • HNF1A
  • TCF1
Protein Name & Synonyms:
Hepatocyte nuclear factor 1-alpha (HNF-1-alpha)(HNF-1A), Liver-specific transcription factor LF-B1(LFB1), Transcription factor 1(TCF-1)
Target Species:
  • Human

Assay Format

Specificity:
The olionucleotide/antibody pair provided in this kit recognizes human HNF-1-alpha in whole lysates and nuclear extracts.
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Lysates
  • Nuclear Extracts
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
  • Semi-Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Introduction

The set of transcription factors that include hepatocyte nuclear factor 1-alpha (HNF-1-alpha), HNF-1-beta, HNF-3-alpha, -beta, and -gamma, HNF4, and HNF6 is preferentially expressed in hepatocytes and plays an important role in liver-specific transcription. HNF-1-alpha was first identified as a DNA-binding protein capable of interacting with the promoters of several hepatic genes, including albumin, alpha1-antitrypsin, and beta-fibrinogen. HNF-1-alpha is composed of three domains, an amino-terminal dimerization domain, a DNA-binding domain belonging to the homeobox family, and a carboxyl-terminal domain that is essential for transactivation of target promoters. HNF-1-alpha and HNF-1-beta share strong homologies in both the dimerization domain and the DNA-binding domain which enable the two proteins to form heterodimers and bind to the same DNA sequences. However, HNF-1-alpha is stronger as a transactivator than is HNF-1-beta. HNF-1-alpha is expressed in polarized epithelia of different organs, including the liver, digestive tract, pancreas, kidney, spleen, thymus, testis, keratinocytes and melanocytes in human skin. HNF-1-alpha is crucial in controlling the postnatal functions of the liver, pancreas, and kidney and involving in various metabolic pathways of other organs through the transcriptional activation of genes that play key roles in phenylalanine catabolism, pancreatic beta-cell glucose-sensing, and renal proximal tubular reabsorption of glucose and several other metabolites. In addition, HNF-1-alpha regulates the expression of acute phase proteins, such as fibrinogen, c-reactive protein, and interleukin 1 receptor, which are involved with inflammation. Moreover, HNF-1-alpha might play a possible tumor suppressor role.

Product Features

  • Specific transcription factor-DNA binding assay
  • Perfect alternative to EMSA
  • Easy to perform in an ELISA format
  • Non-radioactive assay
  • High throughput (96-well plate format)
  • Assay can be completed within 5 hours

Application Notes

Kit Components
  • 96-well Strip Microplate pre-coated with DNA probes
  • DNA Binding Buffer
  • Positive Control Sample
  • Specific Competitor DNA probe
  • Non-specific Competitor DNA probe
  • Assay Reagent
  • DTT
  • Wash Buffer
  • Primary Antibody
  • HRP-conjugated Secondary Antibody
  • Antibody Diluent Buffer
  • TMB One-Step Substrate Reagent
  • Stop Solution
Other Materials Required
  • Distilled or deionized water
  • 100 ml and 1 liter graduated cylinders
  • Tubes to prepare sample dilutions
  • Absorbent paper
  • Precision pipettes to deliver 2 µl to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
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  • Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
  1. Prepare all reagents and samples as instructed in the manual.
  2. Add 100 µl of sample or positive control to each well.
  3. Incubate 2 h at RT or O/N at 4 °C.
  4. Add 100 µl of prepared primary antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared HRP-secondary antibody to each well.
  7. Incubate 1 h at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Typical Data

Figure 1Transcription factor assay of HNF-1-alpha from nuclear extracts of HepG2 cells or HepG2 cells treated with IL-6 with RayBio HNF-1-alpha TF Activity Assay Kit.

Transcription factor assay of HNF-1-alpha from nuclear extracts of HepG2 cells or HepG2 cells treated with IL-6 with RayBio HNF-1-alpha TF Activity Assay Kit

Figure 2Transcription factor assay of HNF-1-alpha from nuclear extracts of HepG2 cells or HepG2 cells treated with IL-6 with the specific competitor or non-specific competitor. The result shows specific binding of HNF-1-alpha to the HNF-1-alpha DNA binding site.

Transcription factor assay of HNF-1-alpha from nuclear extracts of HepG2 cells or HepG2 cells treated with IL-6 with the specific competitor or non-specific competitor. The result shows specific binding of HNF-1-alpha to the HNF-1-alpha DNA binding site.

Storage/Stability

Upon receipt, the positive control should be removed and stored at -20° or -80°C. The remainder of the kit can be stored for up to 6 months at 2-8°C from the date of shipment. Opened Microplate Wells or reagents may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge. Note: The kit can be used within one year if the whole kit is stored at -20°C upon receipt. Avoid repeated freeze-thaw cycles.

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品牌介绍

RayBiotech

美国RayBiotech公司于2001年正式成立,由国际公认的蛋白芯片专家黄若磐教授一手创办,总部设在美国亚特兰大科技园。是世界上第一家专业从事蛋白芯片研发和生产的公司。


RayBiotech是一家领先的生命科学公司,提供蛋白质组学发现工具。RayBiotech拥有近150名员工,其目标是向RayBiotech客户(包括来自41个国家的工业界,学术界和研究机构的科学家)提供全球一流的产品和服务。RayBiotech致力于通过生命科学领域的创新和领导地位来加速客户成功。


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