Insulin ELISA 试剂盒
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北京 101111
Quick Overview for Insulin ELISA 试剂盒 (ABIN8069779)
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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 oxidated Insulin (INS) and total Insulin in human cell and tissue lysates, enabling precise assessment of Insulin oxidation 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-Insulin Antibody
- Positive Control
- HRP-Conjugated Streptavidin
- 200 μL
- Wash Buffer
- Assay Diluent
- Lysis Buffer
- TMB One-Step Substrate Reagent
- Stop Solution
- Oxidation Buffer A
- Oxidation Buffer B
- Oxidation Labeling Reagent
- Oxidation Stabilizing Reagent
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试剂未包括
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- 20 % (v/v) trichloroacetic acid (TCA) solution, ice-cold
- 1:1 (v/v) ethanol:ethyl acetate
- Benchtop centrifuge and microcentrifuge (4 °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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Insulin (INS) ELISA Kit
INS 适用: 大鼠 Colorimetric Competition ELISA 123.5 pg/mL - 10000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Insulin (INS) ELISA KitINS 适用: 人 Colorimetric Competition ELISA 61.7 pg/mL - 5000 pg/mL Cell Culture Supernatant, Plasma, Serum
Insulin (INS) ELISA KitINS 适用: 小鼠 Colorimetric Competition ELISA 123.5 pg/mL - 10000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Insulin (INS) ELISA KitINS 适用: 小鼠 Colorimetric Sandwich ELISA 15.6 nlU/mL - 1000 nlU/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Insulin (INS) ELISA KitINS 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 nlU/mL - 1000 nlU/mL Cell Culture Supernatant, Plasma, Serum
Insulin (INS) ELISA KitINS 适用: 大鼠 Colorimetric Sandwich ELISA 78.125 pg/mL - 5000 pg/mL Plasma, Serum, Tissue Homogenate
Insulin (INS) ELISA KitINS 适用: 小鼠 Colorimetric Sandwich ELISA 78.125 pg/mL - 5000 pg/mL Plasma, Serum, Tissue Homogenate
Insulin (INS) ELISA KitINS 适用: Cow Colorimetric Competition ELISA 123.5 pg/mL - 10000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Insulin (INS) ELISA KitINS 适用: Pig Colorimetric Competition ELISA 61.7 pg/mL - 5000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Insulin (INS) ELISA KitINS 适用: 猴 Colorimetric Competition ELISA 123.5 pg/mL - 10000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, 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.
- Label samples with Oxidation Labeling Reagent
- Add Oxidation Stabilizing Reagent to samples.
- Precipitate proteins with ice-cold 20 % TCA.
- Wash protein pellet twice with 1:1 (v/v) ethanol:ethyl acetate.
- Resuspend protein pellets in Oxidation Buffer B.
- 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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- Insulin (INS)
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别名
- Insulin
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背景
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Gene Symbols: INS
Background: The Oxidated Insulin ELISA Kit uses the "biotin probes" method to biotin label carbonyl groups of oxidized proteins. In this method, the biotin probes first react with carbonyl groups of oxidized proteins to form unstable Schiff bases, which are then further reduced to more stable amines. The biotinylated proteins can then be detected by ELISA. An anti-pan Insulin antibody has been coated onto a 96-well plate. Biotinylated samples are pipetted into the wells, and Insulin 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 oxidated Insulin. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of oxidated Insulin 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): Insulin
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基因ID
- 3630
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UniProt
- P01308
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途径
- NF-kappaB Signaling, RTK signaling, Positive Regulation of Peptide Hormone Secretion, Peptide Hormone Metabolism, Hormone Activity, Carbohydrate Homeostasis, ER-Nucleus Signaling, Regulation of Carbohydrate Metabolic Process, Feeding Behaviour, Autophagy, Negative Regulation of intrinsic apoptotic Signaling, Brown Fat Cell Differentiation, Positive Regulation of fat Cell Differentiation
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