
Antigen Information
- P17275
- 3726
- JUNB
- Human
Assay Format
- HER/ErbB Signaling
- HIF-1 alpha Signaling
- IGF Signaling
- JAK/STAT Signaling
- MAPK Signaling
- p53 Signaling
- PI3K-AKT Signaling
- PKC Signaling
- TGF-beta Signaling
- 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
- 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 assay of jun-B from nuclear extracts of K562 cells or K562 cells treated with PMA (50 ng/ml) for 3 hr. A. Western-blot result of jun-B from cytoplasmic and nuclear fractions. B. Transcription factor assay of jun-B from nuclear fractions with the RayBio® Activity Assay Kit.

Figure 2Transcription factor assay of jun-B 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 jun-B to the conserved binding site detected by using the RayBio® jun-B TF Activity Assay Kit.

Storage/Stability
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颗粒型抗原,除了有细菌、红细胞、螺旋体等天然颗粒型抗原,还有吸附有可溶性抗原的非免疫相关颗粒.
颗粒性抗原光镜下可见,比如细菌性抗原、红细胞抗原等;而可溶性抗原在光镜下不可见,如组织浸出液、细菌毒素、蛋白质分子等。
它们不等同于完全抗原和不完全抗原。完全抗原具有免疫原性和抗原性,而不完全抗原只具有抗原性。完全抗原可以是颗粒性抗原,亦可是可溶性抗原,而不完全抗原一般只能是可溶性抗原,不会是颗粒性抗原。
荚膜是某些细菌在细胞壁外包围的一层粘液性物质,一般由糖和多肽组成,是细菌的一种特殊结构。
作用:
①抗吞噬作用:荚膜因其亲水性及其空间占位、屏障作用,可有效抵抗寄主吞噬细胞的吞噬作用。
②黏附作用:荚膜多糖可使细菌彼此间粘连,也可黏附于组织细胞或无生命物体表面,是引起感染的重要因素,具有荚膜的S-型肺炎链球菌毒力强,有助于肺炎链球菌侵染人体;废水生物处理中的细菌荚膜有生物吸附作用,将废水中的有机物、无机物及胶体吸附在细菌体表面上。
③抗有害物质的损伤作用:处于细菌细胞最外层,荚膜犹如盔甲可有效保护菌体免受或少受多种杀菌、抑菌物质的损伤,如溶菌酶、补体等。
④抗干燥作用:荚膜多糖为高度水合分子,含水量在95%以上,可帮助细菌抵抗干燥对生存的威胁。
⑤当缺乏营养时,荚膜可被利用作碳源和能源,有的荚膜还可作氮源。
所以一个细菌或病毒可以有很多的抗原。

