Stress Response Human Transcription Factors Biology-on-Array

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Biology-on-Array

Stress Response Human Transcription Factors siRNA System: TSPH-004A The Biology-on-Array is an innovative screening tool for identifying the transcription factors that regulate a gene of interest. This system utilizes real-time PCR as an end-point assay coupled with siRNA technology The Human Stress Response Transcription Factors siRNA plate includes pre-designed siRNA pairs (2 designs per gene) for 84 key genes involved in altering gene expression in response to a wide variety of cellular stresses. The plate includes transcription factors that recognize stresses, such as DNA damage, drugs, heat shock, heavy metals, hypoxia, reactive oxygen species, unfolded proteins, UV light, and even viral infection. Transcription factors that mediate the responses to those stress, like apoptosis, cell cycle arrest, and inflammation are also represented by the plate as well as nuclear receptors and other factors responsible for key stress response signaling pathways, such as NFKB and JAK / STAT. The knockdown of each specific transcription modulator results in a unique changes to the transcriptome. The high-throughput first strand kit enables conversion of 96 total RNA samples into cDNA. The real-time PCR assay can be used to analyze expression of any gene in these cDNA samples. The change in the expression of the gene in specific samples(s) reveals that corresponding epigenetic factor(s) are involved in controlling the expression of this gene.

Functional Gene Grouping

Kit Components

How It Works

Manuals & Data Analysis

Gene Table

Apoptosis: AHR, ATF5, CEBPB, DDIT3, E2F1, FOXL2, FOXO1, FOXO3, NFKB1, PPARD, RELA, TP53.

Cell Cycle Regulation: E2F1, E2F4, FOXO4, HIF1A, MYC, RB1, TP53.

DNA Damage: DDIT3, TP53.

Drug Response: AHR, ARNT, DDIT3, PPARD.

Heat Shock Response: AHR, HIF1A, HSF1, HSF2, HSF4, HSFX1.

Heavy Metal Ion Response: MTF1.

Hypoxia: ARNT, EPAS1, HIF1A, USF1, USF2.

Immune Response: CEBPB, PPARG, XBP1.

Inflammation: CEBPB, FOS, HIF1A, NFATC3, NFKB1, RELA.

Nutrient Response: DDIT3, PPARG, RXRA, SREBF1, TP53.

Oncogenes & Proto-Oncogenes: ELF2, ELK1, FOS, JUN, JUND, MAFF, MAX, MYC, RB1, REL, RELA, RELB, TP53, TP63, TP73.

Oxidative Stress: ARNT, DDIT3, HIF1A, TFAM.

Unfolded Protein Response / Endoplasmic Reticulum Stress: ATF4, ATF6, CREB3L3, DDIT3, MSRB2, SREBF1, SREBF2, TP53, XBP1.

UV Stress: USF1.

Viral Infection: IRF3, RELA, STAT1, STAT3.

Activating Transcription Factors: ATF1, ATF2, ATF3, ATF4, ATF5, ATF6.

Forkhead Box Proteins: FOXA2, FOXL2, FOXO1, FOXO3, FOXO4.

Nuclear Receptors: AHR, AR, ARNT, ESR1, ESR2, NR0B1, NR3C1, NR3C2, PGR, PPARD, PPARG, RORA, RXRA, VDR.

Signal Transduction & Cell Signaling:
Calcium & cAMP Signaling: CEBPB, CREB1, CREB3, CREB3L3, NFAT5, NFATC3.
JAK / STAT Signaling: STAT1, STAT3.
NFKB Signaling: NFKB1, REL, RELA, RELB
WNT Signaling: CTNNB1, TCF3.

Others: BACH1, BACH2, EGR1, GTF2I, KLF2, NFE2L2, NFYA, NKX2-5, POU5F1, PAX6, PBX1, RUNX1, RUNX2, SP1, SRF, USF1, USF2.

Functional Gene Grouping

Kit Components

How It Works

Manuals & Data Analysis

Component	Specifications
siRNA plate	Cell Culture-ready Plate. 84 wells contain two siRNAs targeted against a single gene. 12 wells contain control siRNAs (positive, negative and transfection)
96 Sample RNA Isolation Kit	RNeasy® 96 Plates Elution Microtubes CL, racked Caps for Elution Microtubes in strips Buffer RLT Buffer RW1 Buffer RPE RNase-free water
HT First Strand Kit (included in the system)	GE2 (Genomic DNA Elimination Buffer) BC4 (RT Master mix) 96-well PCR Plate 12-well Strip TubesTube Aluminum Foil Sealing Films Compression mat

Storage Conditions

Please check the kit components immediately after you receive this package. SABiosciences is only responsible for missing items reported within two (2) business days of receipt.

The Biology-on-Array Systems are shipped in two boxes (Box 1 on Dry Ice and Box 2 at ambient conditions). Keep the siRNA plate, GE2 and BC4 at -20ºC for long-term storage.
The Systems are stable for six (6) months at the recommended temperature.
Do not use kit beyond the expiration date printed on the label.

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Functional Gene Grouping

Kit Components

How It Works

Manuals & Data Analysis

Overview of Biology-on-Array Protocol.

The Biology-on-Array is comprised of a siRNA plate, a HT first strand kit and data analysis software.

The siRNA plate is a cell culture grade 96-well plate having pre-dispended 84 siRNA pair (2 siRNA design per gene) targeting biologically-relevant groups of transcription regulators (transcription factors or epigenetic regulators) and important RNAi negative and positive controls.
The RNeasy® 96 Kit isolates total RNA from the 96 siRNA-treated samples.
The HT First Strand Kit synthesizes first strand cDNA from the 96 siRNA treated samples while eliminating genomic DNA in approximately 20 minutes.
The Excel-based data analysis software for Biology-on-Array performs the ΔΔCt based fold-change calculations from uploaded raw threshold cycle data from the gene-specific real time-PCR assay. The spreadsheet delivers results in a tabular format and assists in outlier identification and hit selection.

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