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ASB's EASYcis™ promoter--reporter systems are designed for quickly
validation
of GREMA data when multiple promoters were demonstrated for its interaction with
a specific transcription Factor (TF). Two groups of standard vector sets are provided
by ASB for both transient transfection assay and establishing a stable cell lines. | |
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1: Promoter- Luciferase Assay:
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Over 20,000 human basic promoter regions for each unique target
gene have been systematically
analyzed by ASB's team and cloned in Renilla Luciferase based vector. This group of
vector will be
valuable reagents for quickly data validation when a specific TF interaction with
various promoters
has been obtained using GREMA technology. Co-transfection of TF full-length cDNA (make this superlink
to full-length TF cDNA product section). and promoter-luciferase vector will help to rapidly validate
TF/promoter interaction for up- or down regulating of gene transcription. | |
| | A: Customer target gene promoter mapping program: (Human genome Only) | |
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Through genomic-wide human promoter mapping program, ASB has established internal validated human basic
promoter region for more than 20,000 genes. By sending your target information, ASB bioinformatic team can help you map out its
corresponding promoter region using its proprietary 5' mRNA "true extention" program.
Please click here for more information:
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B: Oligo design for PCR validation of ChIP | |
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One of the critical steps in Chromatin immunoprecipitation (ChIP) assay is a validation of TF specific binding promoter DNA
in the pull-down complex using PCR based assay. Due to a high GC containing and rich repetitive elements in human promoter
region, design of oligo for specific amplification of human promoter DNA fragment becomes a challenge job in the field. Currently,
ASB has individually designed and PCR validated more than 20,000 human promoter using its unique in house developed "matrix
selection" algorithm. In order to help our customer, this validated oligo design for each of human gene / promoter could be
accessed by simple send your target accession number and received recommended oligo design in 24 hours in your email. Please
click here for more information:
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| | C: Promoter-ready Vector: | |
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After cloning more than 20,000 human promoters in report vector like Renilla
luciferase, and DNA sequence validated for each of them. You can simple click search bar and access a specific promoter-report
ready plasmids for your project. | |
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| | 2: Stable Cell Line Establishing using a TF specific report plasmid : | |
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Most gene transcription regulations are controlled at multiple interaction of transcription factors with promoter region.
In order to monitor one TF activity at a time such as NFkb in drug screening process, Single TF specific DNA binding site
in the report plasmid will be the best tool in this type of assay.
Add up-stream TF specific DNA binding site on minimal TATA box fragment could convert report gene into a single TF specific
report construct. In aiming to provide customer quickly and convenient creation of a transcription regulation specific stable
cell lines to study in vivo activation of signal transduction pathway, ASB has created a group of constructs which not only
carry a single TF specific responsible report gene, but also selective markers such as HSV TK promoter driven hygromycin or
nemycin phosphotransferase expression cassette for establishing a stable report cell line. The offers fast and efficient
selection of stably transfected cells (with hygromycin or Geneticine®/G418, respectively). The cis-reporting vectors could
also be used in transient assays before committing ones efforts to stable cell lines.
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PRODUCTS LIST: | |
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The EasycisTM cis-reporting system includes a group of vectors covers signal
transduction pathways converging at several common enhancer elements including: | |
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Product
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Size
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Cat. #
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Price
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Contact ASB Sale for quantity pricing.
U.S. Pricing Only. Click here for international sales
Your initial order with ASB may qualify for an introductory discount!
Call for details. | |
| | Kit Contents:
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40µg of transfection-grade report plasmid.
40µg of transfection-grade control plasmid.
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| | References:
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| | The regulatory cis-elements and the firefly (Photinus pyralis) luciferase reporter gene
flanking the TATA box are depicted below, the hygromycin or neomycin expression cassette is also shown.
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Tactic for high sensitivity and specificity in GREMA:
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A cloning vector (pHTS-MCS, Cat# ASB-2130) with convenient cloning sites is also available so
that one can create synthetic promoters to monitor new signaling pathways of interest.
Some of the common features are:
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• Hygromycin or neomycin resistance cassette for efficient stable cell selection
• Ready to use transfection purity plasmid provided
• Unique restriction site for linearization of vector improves stable cell selection efficiency
High copy number plasmid for easy plasmid preparation
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