人Hif1a + BNIP3 Hypoxia In Cell ELISA试剂盒(IR) (ab129733)

概述

  • 产品名称
    人Hif1a + BNIP3 Hypoxia In Cell ELISA试剂盒(IR)
  • 检测方法
    IR
  • 样品类型
    Adherent cells, Suspension cells
  • 检测类型
    Cell-based (quantitative)
  • 实验步骤
    Multiple steps standard assay
  • 种属反应性
    与反应: Human
  • 产品概述

    ab129733 is an In-Cell ELISA (ICE) assay kit that uses quantitative immunocytochemistry to measure HIF1 alpha and BNIP3 protein levels in cultured cells. Cells are fixed in a microplate and targets of interest are detected with highly specific, well-characterized antibodies. Relative protein levels are quantified using IRDye®-labeled Secondary Antibodies and IR imaging using a LI-COR® Odyssey® or Aerius® system.

     

    Hypoxia and the cellular response to hypoxic environment is a central topic in studies of metabolism, cancer progression and development and stem cells. A key player is the transcription factor HIF1 alpha (hypoxia inducible factor 1 alpha) which is stabilized at the protein level in response to decreased oxygen tension. HIF1 alpha then promotes transcription of a number of factors that alters cellular physiology. This Hypoxia ICE assay kit provides duplexed measurements of the transcription factor HIF1 alpha and the HIF1A responsive protein BNIP3.

     

    Plates are available in our ICE (In-Cell ELISA) Support Pack (ab111542) which can be bought seperately.

  • 经测试应用
    适用于: In-Cell ELISAmore details
  • 平台
    Microplate

性能

应用

Our Abpromise guarantee covers the use of ab129733 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
In-Cell ELISA Use at an assay dependent concentration.

图片

  • Sample experiment using ab129733 on HeLa cells treated with a titration of DFO. HeLa cells were seeded to an amine coated 96-well microplate and the following day treated with a titration of DFO. After 24h of DFO exposure, the cells were fixed and stained as described in the protocol and the normalized data is presented here +/- SD (as described in the protocol and data analysis sections). HIF1 alpha results show DFO concentrations ≥10µM induce HIF1A protein levels in a dose dependent manner.
  • Sample experiment using ab129733 on HeLa cells treated with a titration of DFO. HeLa cells were seeded to an amine coated 96-well microplate and the following day treated with a titration of DFO. After 24h of DFO exposure, the cells were fixed and stained as described in the protocol and the normalized data is presented here +/- SD (as described in the protocol and data analysis sections). BNIP3 levels are increased with DFO concentrations ≥10µM.
  • Antibody specificity demonstrated by immunocytochemistry. Primary antibodies used in this assay kit were validated by staining HeLa cells +/- treatment with 1mM DFO (24h) and imaged by fluorescent microscopy. HIF1 alpha staining is absent in untreated cells and induced by DFO treatment. HIF1 alpha localizes to the nucleus (as seen by co-localization with the DNA stain DAPI) as expected.
  • Antibody specificity demonstrated by immunocytochemistry. Primary antibodies used in this assay kit were validated by staining HeLa cells +/- treatment with 1mM DFO (24h) and imaged by fluorescent microscopy. BNIP3 staining is nearly undetectable in untreated HeLa cells but is induced by DFO treatment. BNIP3 appears to has a mitochondrial staining pattern in the DFO-treated samples.
  • Antibody specificity demonstrated by Western Blot. Primary antibodies used in this assay kit were validated by Western Blot using HeLa cell lysates that had been treated with a dose titration of DFO as indicated. (A) The HIF1 alpha band (indicated by arrow) is absent in untreated cells and induced by DFO. (B) Similarly, BNIP3 levels are increased by DFO treatment in a dose-dependent manner.
  • Coefficient of variation for the experiment described in Figure 1.

实验方案

文献

ab129733 has not yet been referenced specifically in any publications.

客户评价及客户问答

There are currently no Customer reviews or Questions for ab129733.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

注册