Product Name Catalog # Price   Qty
Human NFκB-Regulated cDNA Profiling in Cell Lysates AP-3101 $489
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Characterization

Description:

NFkB is a ubiquitous transcription factor that plays a key role in cellular responses to stimuli such as stress, cytokines, free radicals, ultraviolet irradiation, oxidized LDL, and bacterial or viral antigens. Incorrect regulation of NFkB has been linked to cancer, inflammatory and autoimmune diseases, septic shock, viral infection, and improper immune development. When NFkB is activated, it is dissociated from its inhibitor IkB and moves from the cytoplasm to the nucleus, where it binds to target DNA elements and positively regulates the transcription of genes involved in immune and inflammatory responses, cell growth control, and apoptosis. To get a better overview of gene expression of NFkB regulated genes, Signosis has developed a cDNA Plate Array which couples its CLTM cDNA synthesis kit which allows reverse transcription directly from cell lysates without RNA preparation. This plate-based assays allows profiling the expression of 20+ NFkB-regulated genes directly in cell lysates. This assay provides quantitative data, allowing researchers to screen multiple samples, as well as high sensitivity, allowing researchers to use less sample. 


Applicable Grid:

List of Applicable Genes

BCL2 IL6  Myc  BCL2 IL6  Myc  BCL2 IL6  Myc  BCL2 IL6  Myc 
Beta-actin  IL8  NOS2  Beta-actin  IL8  NOS2  Beta-actin  IL8  NOS2  Beta-actin  IL8  NOS2 
CCND1 GAPDH  p53 CCND1 GAPDH  p53 CCND1 GAPDH  p53 CCND1 GAPDH  p53
Cox-2  IFNg TNFA  Cox-2  IFNg TNFA  Cox-2  IFNg TNFA  Cox-2  IFNg TNFA 
FAS-L  IRF1  TNFR  FAS-L  IRF1  TNFR  FAS-L  IRF1  TNFR  FAS-L  IRF1  TNFR 
IFNB1  MDR1 VCAM1 IFNB1  MDR1 VCAM1 IFNB1  MDR1 VCAM1 IFNB1  MDR1 VCAM1
IL1A  MMP1  VEGFC IL1A  MMP1  VEGFC IL1A  MMP1  VEGFC IL1A  MMP1  VEGFC
IL2 Myb Blank IL2 Myb Blank IL2 Myb Blank IL2 Myb Blank

 


Principle

Unlike conventional reverse transcription which needs total RNA, Signosis has developed a CL(TM) cDNA synthesis-based plate array. It can synthesize cDNA probes through reverse transcription directly in cell lysates without RNA preparation. The synthesized cDNA probes are then applied for a plate array hybridization. Targeted genes are specifically captured onto individual wells on a plate through a pre-coated gene-specific oligonucleotide. The captured cDNAs are further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity.

 

 

Data


Human NFkB-regulated cDNA plate array analysis.  HeLa cells were treated with (red bars) or without (light blue) 10ng/ml TNF for 30 min.  Cells were collected for profiling 23 genes with the cDNA plate array assay (top). Time course of IL-6 induction to TNFa treatment in HeLa cells was monitored with the cDNA plate array assay.

Literature

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