NAAA ELISA 试剂盒
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Quick Overview for NAAA ELISA 试剂盒 (ABIN8070165)
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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 s-acylated ASAHL (NAAA) and total ASAHL in human cell and tissue lysates, enabling precise assessment of ASAHL s-acylation 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-ASAHL Antibody
- Positive Control
- HRP-Conjugated Streptavidin
- 200 μL
- Wash Buffer
- Assay Diluent
- Lysis Buffer
- TMB One-Step Substrate Reagent
- Stop Solution
- S-Acylation Buffer A
- S-Acylation Buffer B
- S-Acylation Buffer C
- S-Acylation Buffer D
- S-Acylation Blocking Reagent
- S-Acylation Reduction Reagent
- S-Acylation 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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N-Acylethanolamine Acid Amidase (NAAA) ELISA Kit
NAAA 适用: 人 Colorimetric Sandwich ELISA 0.31 ng/mL - 20 ng/mL Cell Lysate, Tissue Homogenate
N-Acylethanolamine Acid Amidase (NAAA) ELISA KitNAAA 适用: 人 AA 1-359 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Plasma (EDTA), Plasma (citrate), Plasma (heparin), Serum
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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-Acylation 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 Buffers and adding 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-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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- NAAA (N-Acylethanolamine Acid Amidase (NAAA))
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别名
- ASAHL
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背景
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Gene Symbols: NAAA, ASAHL, PLT
Background: The S-Acylated ASAHL ELISA Kit uses a modified "biotin-switch" method to allow for the direct visualization of human S-Acylated ASAHL by ELISA. In this S-Acylation biotin switch assay, unmodified free cysteines are first blocked. S-Acylated cysteines are then selectively reduced for specific labeling with biotin-maleimide reagents, which irreversibly bind to the cysteine thiol that was S-Acylated. Biotinylation of the newly formed thiol groups can then be detected by ELISA. An anti-pan ASAHL antibody has been coated onto a 96-well plate. Biotinylated samples are pipetted into the wells, and ASAHL 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-Acylated ASAHL. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of S-Acylated ASAHL 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): N-acylethanolamine-hydrolyzing acid amidase, Acid ceramidase-like protein, Acylsphingosine deacylase NAAA, N-acylsphingosine amidohydrolase-like (ASAH-like protein)
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基因ID
- 27163
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UniProt
- Q02083
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