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Human NF-κB p52 Activity Assay Kit
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原料试剂技术服务 动物实验室研发实验室
价格
25008
品牌 瑞博奥/RayBio
地区 -
货号 TFEH-p52-1
产地 进口
选择规格
1 Plate Kit
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RayBio多因子检测
瑞博奥(广州)生物科技股份有限公司
中国广东省广州市
营业执照已审核
RayBio是一家总部位于美国乔治亚州的生物技术公司,专注于开发和制造高质量的蛋白质检测工具以及提供相应的服务,聚焦于蛋白质组学的检测分析领域和多组学的应用,以严格的质量控制和创新的技术为基础。 Raybio具有可用于生命科学研究、疾病研究及疾病治疗相关药物、医学技术等功能探究的全线产品,包括国际领先、最多专利、最多指标的抗体芯片、流式多因子芯片等多因子检测试剂盒超700余种;单靶标检的ELISA试剂盒,可用于细胞因子含量、药物作用等检测;以及超过18,000种抗体、小分子产品,可用于WB、ELISA、IF等免疫学实验。 我们的产品和服务被全球的学术、政府和制药实验室广泛使用,用于疾病诊断、药物筛选和基础研究,已助力全球客户在Nature, Nature Medicine, Cell, Lancet, PNAS等世界顶级期刊上发表了数万篇文章。
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产品规格 图文详情 技术文档
产品规格
品牌名称
瑞博奥/RayBio
货号
TFEH-p52-1
国产/进口
进口
规格
1 Plate Kit
图文详情

Introduction

The non-canonical NF-κB pathway is an important arm of NF-κB signaling that predominantly functions through activation of the p52/RelB NF-κB complex. This pathway regulates important biological functions, such as lymphoid organogenesis, B-cell survival and maturation, dendritic cell activation, and bone metabolism. Moreover, deregulated non-canonical NF-κB signaling is associated with lymphoid malignancies. In contrast to the canonical NF-κB pathway, in which NF-κB is inactive and retained in the cytoplasm by its inhibitor, IκB, and only transported to the nucleus through modification and degradation of IκB upon stimulation of various factors, the non-canonical pathway depends on the inducible processing of p100, that functions as both the precursor of p52 and a RelB-specific inhibitor. When cells are stimulated by of a subset of TNF receptor family members, the downstream kinase IκB kinase-α (IKKα) is activated and triggers p100 phosphorylation and processing, which generates p52 and results in RelB/p52 NF-κB complex translocation to the nucleus where it regulates the corresponding downstream genes. It is increasingly clear that this pathway of NF-κB activation differs significantly from the canonical NF-κB pathway in its signaling mechanisms. Therefore, better understanding of the mechanism regulating non-canonical NF-κB activation has important therapeutic values.

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
  • 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 assay of NF-κB p52 from nuclear extracts of HL60 cells or HeLa cells with the RayBio®NF-κBp52 TF Activity Assay.

Transcription factor assay of NF-κB p52 from nuclear extracts of HL60 cells or HeLa cells with the RayBio NF-κBp52 TF Activity Assay.

Figure 2
Transcription factor assay of NF-κB p52 from nuclear extracts of HL60 cells or HeLa cells with the specific competitor or non-specific competitor. The result shows specific binding of NF-κB p52 to the NF-κB non-canonical DNA binding site.

Transcription factor assay of NF-κB p52 from nuclear extracts of HL60 cells or HeLa cells with the specific competitor or non-specific competitor. The result shows specific binding of NF-κB p52 to the NF-κB non-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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