U-Blot® Human Hydroxylated-HIF-1 alpha (P402) and HIF-1 alpha ELISA Kit

The Hydroxylated-HIF-1alpha (Pro402) and HIF-1alpha 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 and 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 or biotinylated anti-HIF-1alpha antibody is used to detect HIF-1alpha. 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 hydroxylated HIF-1alpha (Pro402) or HIF-1alpha bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.Notably, this kit allows simultaneous measurement of methylated protein and pan protein in a single experiment for accurate data normalization. It supports high-throughput screening of numerous different cell lysates without the time-consuming and labor-intensive Western Blot analysis. Featuring minimal hands-on operation time, simple workflow, and a completely non-radioactive detection system, this assay provides a convenient and safe solution for histone methylation research.