RayBiotech/人NF-KappaB p65转录因子活性测定/1平板试剂盒/TFEH-p65-1
商品编号:
TFEH-p65-1
品牌:
RayBiotech
市场价:
¥0.00
美元价:
0.00
产品分类:
细胞计数仪
公司分类:
Cell_counter
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:
- Q04206
Gene ID:
- 5970
Gene Symbols:
- NFKB3
- RELA
Protein Name & Synonyms:
Transcription factor p65 (Nuclear factor NF-kappa-B p65 subunit) (Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3)
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 p65 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 < li="">
- Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared HRP-secondary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Typical Data
Figure 1 Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells or HeLa cells treated with TNF-α. After stimulation with TNF-α, activated NF-κB p65 is translocated into the nucleus where it binds with its corresponding DNA.
Figure 2 Transcription factor activity assay of NF-κB p65 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 p65 to the NF-κB DNA binding site.
Figure 3 Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells treated with TNF-α and purified NF-κB p50. No cross-reactivity has been observed between NF-κB p65 and p50.
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.
No posts found
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。
联络我们