Antigen Information
- Q9H9S0
- 79923
- NANOG
- Human
Assay Format
- Human
- Cell Lysates
- Nuclear Extracts
- Sandwich-based
- Semi-Quantitative
Product Specifications
Introduction
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
- 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
- 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
- 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 1Transcription factor activity assay of Nanog from nuclear extracts of P19 cells with the RayBio® Nanog TF-Activity Assay Kit.
Figure 2Transcription factor activity assay of Nanog from nuclear extracts of P19 cells with the specific competitor or non-specific competitor. The result shows specific binding of Nanog to the conserved DNA binding site.
Storage/Stability
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如题,试剂商推荐这家的,然后网上查了一下就是武汉三鹰啊,国产的,有点虚啊?请前辈指点迷津啊
利用CD3/CD4/IL-17的抗体组合(不同荧光素标记),具体步骤可按照淋巴细胞TH1/2亚型分析的protocol!
一般都是先做CD4的surface staining
然后再做IFN-gamma和IL-17的intrecellular staining
测流式后不用水洗
不用问为什么?_?
如果一个抗体可以用来“做western blot,免疫荧光,免疫共沉淀,ELISA”,我估计着是HRP或碱性磷酸酶或生物素之类标记的,然后需要再加荧光标记的二抗,这类抗体也是不能做流式用的。
我只用过流式的抗体来做免疫荧光(直接染色),效果还不错,但是反过来就没试过。
楼上两位还是要仔细看下产品说明书
尽量使用已经用荧光素标记好的单克隆抗体。
如果是单色实验,荧光素的亮度越强越好,比如标记了APC的抗体就要比标记了pacific blue的在相同抗原量,抗体用量相同的情况下,要亮很多
如果是多色实验,除了不要使用光谱重叠度高的荧光素以外,还要搭配染色指数。简单的说,就是用亮度强的荧光素标记的抗体来染水平低的抗原,而用较弱的荧光素标记的抗体来染高表达的抗原。
暂无品牌问答