Procarta TF Plex Assay

Procarta TF Plex Assay

 

Transcription factors (TFs), which play crucial roles in the regulation of gene expression in the human genome, are highly regulated by a variety of mechanisms. A single extracellular stimulus can trigger multiple signaling pathways, and these in turn can activate multiple TFs to mediate the inducible expression of target genes.

Alterations in the activities of TFs are often associated with human diseases, such as altered activating factor 1, estrogen receptor, and p53 function in cancer, nuclear factor kB in inflammatory diseases, and peroxisome proliferator-activated receptor gamma in obesity.

A systematic assay for profiling the activation of TFs will aid in elucidating the mechanisms of TF activation, reveal altered TFs associated with human diseases, and aid in developing assays for drug discovery.

 

Panomics has developed a 40-plex fluorescent microsphere-based TF activation assay system with a 96-well plate format. The assay system enables high-throughput profiling of the DNA binding activity of TFs in multiple samples with high sensitivity.

TF Plex Panel 1

Panomics offers 40 different Transcription factors for Panel 1. Users can choose from the 40 TF's below to create their own Plex set.

 

TF Plex Panel 2

Panomics offers 43 different Transcription factors for Panel 1. Users can choose from the 43 TF's below to create their own Plex set.

 

Monitoring PMA-Mediated Induction in Nuclear Extracts

 

Panomics Procarta Transcription Factor Assay was used to simultaneously measure the activity of 40 TFs in a single assay well.

HeLa cells were serum starved overnight and subsequently stimulated PMA or a vehicle control for 4 hours. As expected, PMA induced AP1 and NF-E2 and the activity of NF-1 and STAT4 were unchanged. Sample inductions from the Procarta TF assay were confirmed by EMSA Gel Shift assays.

Lane 1 = Free Probe.

Lane 2 = HeLa Nuclear Extract.

Lane 3 = PMA Induced HeLa Nuclear Extract.

 

Procarta TF Plex Kits (Panel 1 and Panel 2) are available in ¼- and 1-plate 96-well formats for our off-the-shelf 40 or 43 Plex kits.

All TF Plex Kits including "By Request" kits contain all the reagents required to detect the presence of activated transcription factors for analysis on the Luminex system. Every kit also includes reagents for the preparation of nuclear extracts from adherent and non-adherent cells.

 

By Request Procarta TF Assay Kits:

 

By Request (customer specified) 3-42 plex Procarta TF assays are available in 1-plate formats. Customer selected TFs from our Procarta TF panel are provided in a premixed ready-to-use format.

All By Request Procarta TF Assay Kits must include NFkB in order to run the positive and negative controls provided in the kit. For example, if you require 2 assays such as AP-1 and PPAR, you will need to choose PC5900 for a 3 plex, 1plate assay that will inlcude NFkB, AP-1 and PPAR

 

 

Extraction Kits for Procarta TF Plex Assay

 

The Nuclear Extraction Kit contains reagents and procedures for the preparation of nuclear extracts for use in our Procarta TF Assays. The Procarta TF Nuclear Extraction Kit contains sufficient reagents for the preparation of 40 nuclear extracts from cultured cells grown in 6-well culture plates or 20 nuclear extracts from cultured cells grown in 100-mm culture dishes.

 

The Procarta TF Whole Cell Lysis Kit contains reagents and procedures for the preparation of whole cell lysates for use in our Procarta TF Assays. The Procarta TF Whole Cell Lysis Kit contains sufficient reagents for the preparation of 40 whole cell lysates from cultured cells grown in 6-well culture plates or 240 whole cell lysates from cultured cells grown in 96-well culture plates.