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Human Phospho-IGF1R (Y1165/1166) and Total IGF1R
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原料试剂技术服务 动物实验室研发实验室
价格
21072
品牌 瑞博奥/RayBio
地区 -
货号 PEL-IGF1R-Y1165-T-2
产地 进口
选择规格
2 x 96-Well Strip Microplate Kit
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RayBio多因子检测
瑞博奥(广州)生物科技股份有限公司
中国广东省广州市
营业执照已审核
RayBio是一家总部位于美国乔治亚州的生物技术公司,专注于开发和制造高质量的蛋白质检测工具以及提供相应的服务,聚焦于蛋白质组学的检测分析领域和多组学的应用,以严格的质量控制和创新的技术为基础。 Raybio具有可用于生命科学研究、疾病研究及疾病治疗相关药物、医学技术等功能探究的全线产品,包括国际领先、最多专利、最多指标的抗体芯片、流式多因子芯片等多因子检测试剂盒超700余种;单靶标检的ELISA试剂盒,可用于细胞因子含量、药物作用等检测;以及超过18,000种抗体、小分子产品,可用于WB、ELISA、IF等免疫学实验。 我们的产品和服务被全球的学术、政府和制药实验室广泛使用,用于疾病诊断、药物筛选和基础研究,已助力全球客户在Nature, Nature Medicine, Cell, Lancet, PNAS等世界顶级期刊上发表了数万篇文章。
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产品规格 图文详情 技术文档
产品规格
品牌名称
瑞博奥/RayBio
货号
PEL-IGF1R-Y1165-T-2
国产/进口
进口
规格
2 x 96-Well Strip Microplate Kit
图文详情

Introduction

RayBio® Phospho-IGF1R (Tyr1165/1166) and Total IGF1R ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated IGF1R protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-IGF1R and total IGF1R. An anti-pan IGF1R antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and IGF1R present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-IGF1R (Tyr1165/1166) antibody is used to detect phosphorylated IGF1R or biotinylated anti-IGF1R antibody is used to detect pan IGF1R. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or 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 IGF1R (Tyr1165/1166) or pan IGF1R bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Product Features

  • Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
  • Screen numerous different cell lysates without performing a Western Blot analysis
  • Minimal hands-on time, convenient, and non-radioactive material

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Anti-Phospho Antibody
  • Anti-Pan Antibody
  • HRP-Conjugated Secondary Antibody
  • Streptavidin-Conjugated HRP
  • Assay Diluent
  • TMB One-Step Substrate
  • Stop Solution
  • Lysis Buffer
  • Positive Control Sample
Other Materials Required
  • Distilled or deionized water
  • 100 ml and 1 liter graduated cylinders
  • Tubes to prepare sample dilutions
  • Protease and Phosphatase inhibitors
  • Precision pipettes to deliver 2 µl to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
  • Benchtop rocker or shaker
  • Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
  1. Prepare all reagents and samples as instructed in the manual.
  2. Add 100 µl of sample or positive control to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 µl of prepared primary antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.
  7. Incubate 1 h at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Typical Data

Positive Control
Jurkat cells were treated with PV (Pervanadate). Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
IGF Stimulation of HepG2 Cell Line
HepG2 cells were treated or untreated with IGF. Cell lysates were analyzed using this phosphoELISA and Western Blot.

Storage/Stability

Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at –20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C.
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