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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 c-Jun Transcription Factor Activity Assay/1 Plate Kit/TFEH-CJUN-1
产品编号:TFEH-CJUN-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 c-Jun Transcription Factor Activity Assay/1 Plate Kit/TFEH-CJUN-1
商品介绍

Antigen Information

Accession Number:
  • P05412
Gene ID:
  • 3725
Gene Symbols:
  • JUN
Protein Name & Synonyms:
Transcription factor AP-1, Activator protein 1 (AP1), Proto-oncogene c-Jun, V-jun avian sarcoma virus 17 oncogene homolog, p39
Target Species:
  • Human
Target Name:
c-Jun

Assay Format

Specificity:
The olionucleotide/antibody pair provided in this kit recognizes mouse c-Jun 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

Activator protein-1 (AP-1) is a sequence-specific transcriptional activator composed of members of the Jun (c-Jun, JunB, and JunD) and Fos (c-Fos, FosB, Fra1, and Fra2) families in formats of homo- and heterodimers. These proteins belong to the bZIP group of DNA binding proteins with the ability to bind a common consensus sequence-defined AP-1-binding site. Jun and Fos proteins can also dimerize other basic leucine zipper proteins such as ATF, CCAAT enhancer-binding protein, Maf, and NF-E2. Jun-Jun and Jun-Fos dimers bind preferentially to TPA responsive element (TRE), whose consensus is TGAGTCA, whereas Jun-ATF dimers prefer to bind to the c-AMP-responsive element (CRE) whose consensus is TGAGCTCA. Inside cells, AP-1 activity is induced by an incredible diversity of signals, including growth factors, cellular stress, ionizing and ultraviolet irradiation, DNA damage, oxidative stress, neuronal depolarization antigen binding by T or B lymphocytes, and cytokines. The mechanisms involved in the induction of AP-1 activity are mediated through changes in the expression of AP-1 components or post-translational modification or both, resulting in regulation of their trans-activity positively or negatively. For example, stimulation by growth factors or by activating mutations in cytoplasmic effectors such as ras and raf, results in AP-1 activation by triggering the ERK signaling pathway. On the other hand, AP-1 responses to proinflammatory cytokines and UV radiation mostly depend on two other MAPK cascades, JNK and p38. The result is that AP-1 regulates various target genes executing biological functions such as cell proliferation, differentiation, apoptosis, or cell death.

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 c-JUN from nuclear extracts of K562 cells or K562 cells treated with PMA (50 ng/ml) for 3 hr. A. Western blot of c-Jun from cytoplasmic and nuclear fractions. B. Detection of c-Jun from nuclear fractions with the RayBio® Activity Assay Kit.

Transcription factor assay of c-JUN from nuclear extracts of K562 cells or K562 cells treated with PMA (50 ng/ml) for 3 hr.

Figure 2Transcription factor assay of c-Jun from nuclear extracts of K562 cells or K562 cells treated with PMA (50 ng/ml) for 3 hr with the specific competitor or non-specific competitor. The result shows specific binding of c-Jun to the conserved binding site detected by using the RayBio® c-Jun TF Activity Assay Kit.

Transcription factor assay of c-Jun from nuclear extracts of K562 cells or K562 cells treated with PMA (50 ng/ml) for 3 hr with the specific competitor or non-specific competitor.

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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