Human Hydroxylated-HIF-1alpha (P402) ELISA Kit

掲載日情報:2017/06/21 現在Webページ番号:204170

RayBiotech社のHuman Hydroxylated-HIF-1alpha (P402) ELISA Kitです。
本製品は研究用です。研究用以外には使用できません。

RayBiotech社ELISA kit

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Human Hydroxylated-HIF-1alpha (P402) ELISA (1 x 96-Well Strip Microplate Kit)
10日程度 ※ 表示されている納期は弊社に在庫がなく、取り寄せた場合の目安納期となります。 0
Human Hydroxylated-HIF-1alpha (P402) ELISA (96-Well Strip Microplate Kit)
10日程度 ※ 表示されている納期は弊社に在庫がなく、取り寄せた場合の目安納期となります。 0
Human Hydroxylated-HIF-1alpha (P402) ELISA (96-Well Strip Microplate Kit)
10日程度 ※ 表示されている納期は弊社に在庫がなく、取り寄せた場合の目安納期となります。 0

[在庫・価格 :2024年03月29日 00時00分現在]

※ 表示されている納期は弊社に在庫が無く、取り寄せた場合の納期目安となります。

Human Hydroxylated-HIF-1alpha (P402) ELISA (1 x 96-Well Strip Microplate Kit)

文献数: 0

Human Hydroxylated-HIF-1alpha (P402) ELISA (96-Well Strip Microplate Kit)

文献数: 0

Human Hydroxylated-HIF-1alpha (P402) ELISA (96-Well Strip Microplate Kit)

文献数: 0

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製品説明

■Introduction

RayBio® Hydroxylated-HIF-1alpha (Pro402) 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 hydroxylated HIF-1alpha 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 hydroxylated-HIF-1alpha. An anti-pan HIF-1alpha antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and HIF-1alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-Hydroxylated-HIF-1alpha (Pro402) antibody is used to detect hydroxylated HIF-1alpha. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG 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 hydroxylated HIF-1alpha (Pro402) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

■Product Features

  • Rapidly measure Hydroxylated protein in lysates
  • Screen numerous different cell lysates without performing a Western Blot analysis
  • Minimal hands-on time, convenient, and non-radioactive material

■Specificity

The antibody pair provided in this kit recognizes human Hydroxylated-HIF-1alpha hydroxylated at site Proline-402

■Target Protein Information

Synonyms: Hypoxia-inducible factor 1-alpha (HIF-1-alpha) (HIF1-alpha) (ARNT-interacting protein) (Basic-helix-loop-helix-PAS protein MOP1) (Class E basic helix-loop-helix protein 78) (bHLHe78) (Member of PAS protein 1) (PAS domain-containing protein 8)

Accession Number: Q16665

Gene Names: HIF1A BHLHE78 MOP1 PASD8

Gene ID: 3091

■Species Detected

Human

■Application Notes

Kit Components

  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Anti-Hydroxylated Antibody
  • HRP-Conjugated Secondary Antibody
  • 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

HeLa cells were treated with MG132. 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.

MG132 and DMOG Inhibition of HeLa Cell Line

HeLa cells were treated or untreated with MG132 and DMOG. Cell lysates were analyzed using this ELISA 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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製品詳細

■ Antigen Information

Gene Symbol:

  • HIF1A

■ Product Specifications

Size: 1, 2, or 5 x 96-Well Strip Microplate Kit

■ Array/ELISA Format

Pathway:

  • Apoptosis

Species Detected:

  • Human

Solid Support: 96-well Microplate

Design Principle:

  • Sandwich-based

Quantitative/Semi-Quantitative: Semi-Quantitative

Method of Detection: Colorimetric

Compatible Sample Types:

  • Cell Lysates
  • Tissue Lysates

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お問い合わせ先

(テクニカルサポート 試薬担当)

reagent@funakoshi.co.jp

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