NEDD8 ELISA 试剂盒
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Quick Overview for NEDD8 ELISA 试剂盒 (ABIN8070200)
抗原
See all NEDD8 ELISA试剂盒抗原表位
适用
检测方法
实验类型
应用范围
样品类型
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原理
- Measures ubiquitinated NEDD8 and total NEDD8 in human cell and tissue lysates, enabling precise assessment of NEDD8 ubiquitination by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.
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Analytical Method
- Quantitative
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组件
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- Size / Qty
- Microplate Coated With Anti-Pan-NEDD8 Antibody
- Anti-Ubiquitination Detection Antibody
- Anti-NEDD8 Detection Antibody
- HRP-Conjugated Streptavidin
- 200 μL
- Wash Buffer
- Assay Diluent
- Lysis Buffer
- TMB One-Step Substrate Reagent
- Stop Solution
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试剂未包括
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- Microplate reader capable of measuring absorbance at 450 nm
- Protease and Phosphatase inhibitors
- Precision pipettes to deliver 2 μL to 1 mL volumes
- Adjustable 1-25 mL pipettes for reagent preparation
- 100 mL and 1 liter graduated cylinders
- Log-log graph paper or computer and software for ELISA data analysis
- Absorbent paper
- Tubes to prepare sample dilutions
- Distilled or deionized water
- Shaker
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应用备注
- Optimal working dilution should be determined by the investigator.
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板类型
- Pre-coated
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实验流程
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- Prepare all reagents and samples as instructed.
- Add 100 μL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
- Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
- Add 100 μL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
- Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
- Add 50 μL Stop Solution to each well. Read at 450 nm immediately.
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限制
- 仅限研究用
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储存条件
- -20 °C
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储存方法
- The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.
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- NEDD8 (Neural Precursor Cell Expressed, Developmentally Down-Regulated 8 (NEDD8))
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别名
- NEDD8
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背景
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Gene Symbols: NEDD8
Background: The Ubiquitinated-NEDD8 and Total-NEDD8 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human ubiquitinated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan NEDD8 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and NEDD8 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-ubiquitination antibody is used to detect ubiquitinated NEDD8 or biotinylated anti-NEDD8 antibody is used to detect total NEDD8 . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ubiquitinated NEDD8 or total NEDD8 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Protein Name (Synonyms): Ubiquitin-like protein NEDD8, Neddylin, Neural precursor cell expressed developmentally down-regulated protein 8 (NEDD-8)
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基因ID
- 4738
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UniProt
- Q15843
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途径
- Ubiquitin Proteasome Pathway
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