NF-kB p65 ELISA 试剂盒
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Quick Overview for NF-kB p65 ELISA 试剂盒 (ABIN8070289)
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See all NF-kB p65 (NFkBP65) 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
- S-Nitrosylated NFkB ELISA Kit uses a modified 'biotin-switch' method to allow for the direct visualization of human, rat, canine (dog), frog, and rabbit S-Nitrosylated NFkB by ELISA application.
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Analytical Method
- Quantitative
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组件
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- Size / Qty
- Anti-Pan-NFKBp65 Microplate
- 96 wells
- Positive Control
- HRP-Streptavidin
- 200 μL
- Wash Buffer
- Assay Diluent B
- Lysis Buffer
- TMB One-Step Substrate Reagent
- Stop Solution
- S-Nitrosylation Buffer A
- S-Nitrosylation Buffer B
- S-Nitrosylation Buffer C
- S-Nitrosylation Buffer D
- S-Nitrosylation Buffer E
- S-Nitrosylation Blocking Reagent
- S-Nitrosylation Reduction Reagent I
- S-Nitrosylation Reduction Reagent II
- S-Nitrosylation Labeling Reagent
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试剂未包括
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- Acetone (pre-chilled (-20 °C))
- 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 positive control or sample dilutions
- Distilled or deionized water
- Shaker
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Nuclear Factor-kB p65 (NFkBP65) ELISA Kit
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Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 人 Colorimetric Sandwich ELISA 0.16 ng/mL - 10 ng/mL Cell Culture Supernatant, Plasma, Serum
Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 小鼠 Colorimetric Sandwich ELISA 0.31 ng/mL - 20 ng/mL Cell Culture Supernatant, Plasma, Serum
Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 人 Colorimetric DNA-Binding ELISA Cell Lysate, Nuclear Extract
Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 人 Colorimetric Sandwich ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 小鼠 Colorimetric Sandwich ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 大鼠 Colorimetric Sandwich ELISA 78.125 pg/mL - 5000 pg/mL Plasma, Serum, Tissue Homogenate
Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 人 Colorimetric Sandwich ELISA 0.15 ng/mL - 10 ng/mL Cell Lysate, Plasma, Serum, Tissue Homogenate
Nuclear Factor-kB p65 (NFkBP65) ELISA KitNFkBP65 适用: 大鼠 Colorimetric Sandwich ELISA 0.15 ng/mL - 10 ng/mL Cell Lysate, Tissue Homogenate
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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, samples and Positive Control as instructed.
- Block unmodified free cysteines using prepared S-Nitrosylation Blocking Buffer at 50 °C with gentle rocking.
- Precipitate proteins with ice-cold acetone.
- Wash protein pellet twice with cold acetone/water mixture.
- Resuspend protein pellets in Buffer containing Reducing and Labeling Reagents.
- Precipitate proteins with ice-cold acetone.
- Resuspend protein pellets.
- Add 100 μL sample and Positive Control to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
- Add 100 μL prepared HRP-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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- NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))
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别名
- NF-kB P65
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背景
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Gene Symbols: RELA, NFKB3
Background: The S-Nitrosylated NF-KB p65 ELISA Kit uses a modified "biotin-switch" method to allow for the direct visualization of human and mouse S-Nitrosylated NF-KB p65 by ELISA. In this S-Nitrosylation biotin switch assay, unmodified free cysteines are first blocked. S-Nitrosylated cysteines are then selectively reduced for specific labeling with biotin-maleimide reagents, which irreversibly bind to the cysteine thiol that was S-Nitrosylated. Biotinylation of the newly formed thiol groups can then be detected by ELISA. An anti-pan NF-KB p65 antibody has been coated onto a 96-well plate. Biotinylated samples are pipetted into the wells and NF-KB p65 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and HRP-conjugated streptavidin is used to detect S-Nitrosylated NF-KB p65. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of S-Nitrosylated NF-KB p65 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): Transcription factor p65 (Nuclear factor NF-kappa-B p65 subunit) (Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3)
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基因ID
- 5970
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UniProt
- Q04206
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途径
- NF-kappaB Signaling, RTK signaling, TCR Signaling, TLR signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Toll-Like Receptors Cascades, S100 Proteins
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