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当前位置: 首页 > 产品中心 > Others > RayBiotech/人NF-KappaB p50转录因子活性测定/1平板试剂盒/TFEH-p50-1
商品详细RayBiotech/人NF-KappaB p50转录因子活性测定/1平板试剂盒/TFEH-p50-1
RayBiotech/人NF-KappaB p50转录因子活性测定/1平板试剂盒/TFEH-p50-1
RayBiotech/人NF-KappaB p50转录因子活性测定/1平板试剂盒/TFEH-p50-1
商品编号: TFEH-p50-1
品牌: RayBiotech
市场价: ¥0.00
美元价: 0.00
产地: 美国(厂家直采)
公司:
产品分类: 其他
公司分类: Others
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍

Antigen Information

Accession Number:
  • P19838
Target Species:
  • Human

Assay Format

Pathway:
  • B Cell Receptor
  • Inflammation
  • NFkB Signaling
  • T Cell Receptor
Specificity:
The olionucleotide/antibody pair provided in this kit recognizes human NF-?B p50 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

NF-κB is a transcription factor that plays a pivotal role in a number of physiological cell processes such as growth, apoptosis, and immune response through regulating the expression of a number of cellular genes. The NF-κB family includes five members: cRel, RelB, p65(RelA), p50(p105, precursor of p50), and p52(p100, precursor of p52), which form homo- or heterodimers when in active status. Normally, NF-κB is inactive and retained in the cytoplasm bound to its inhibitory protein IκB. Upon stimulation of various factors such as infection of pathogens including viruses and bacteria or treatment of cytokines such as IL-1 or TNF-α, IκB proteins are phosphorylated, ubiquitinated and finally degraded. Released NF-κB proteins then form homo- or heterodimers, most commonly p50/p65 or p50/p50, which are transported into the nucleus where they bind to specific DNA sequences of targeted genes and activate transcription. NF-κB also induces expression of its own inhibitor, IκBa, which binds to active NF-κB in the nucleus to terminate the induction of gene expression.

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 1 Transcription factor activity assay of NF-κB p50 from nuclear extracts of HeLa cells or HeLa cells treated with TNF-α. After stimulation with TNF-α for 30 minutes, activated NF-κB p50 is translocated into the nucleus where it binds with its corresponding DNA. A.Western-blot result of NF-κB p50 from cytoplasm and nuclear fractions. B. Transcription factor activity assay of NF-κB p50 from nuclear fractions with the RayBio®NF-κB p50 Transcription Factor-Activity Assay Kit.

Figure 2 Transcription factor activity assay of NF-κB p50 from nuclear extracts of HeLa cells or HeLa cells treated with TNF-α with the specific competitor or non-specific competitor. The result shows specific binding of NF-κB p50 to the NF-κB canonical 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致力于开发、研制、生产及销售以抗体芯片、ELISA/EIA、抗原抗体为主的酶免产品。率先开发了高品质的抗体和蛋白质芯片技术平台,蛋白质芯片能够同时有效地分析生物样品中参与炎症、血管生长、细胞生长、细胞凋亡及信号转导等数百种蛋白的表达水平。近400页的产品目录,主要涉及蛋白质芯片、抗体芯片、抗体、免疫试剂、诊断试剂等近万种,拥有先进的技术优势。许多发表在Nature、Nature Medicine、 Cell、 PNAS、Lancet等世界顶级期刊上的文章中均使用了瑞博奥的产品,公司的芯片技术和产品质量都得到了广泛的肯定。