ALEXIS Biochemicals: EGR-3 Protein (human) (recombinant)

ALX-201-075

Revised 02-May-06

EGR-3 Protein (human) (recombinant)
SYNONYMS	Early Growth Response Protein 3 (human) (recombinant) Pilot (human) (recombinant)
PRODUCT LINE	DNA Regulation / Transcription
PRODUCT CATEGORY	Egr Protein Family/Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-075-1		1 Vial	395.00 USD

Product Specification

SOURCE/HOST:	Produced in Sf9 cells.
QUANTITY:	75µl, corresponding to >25 band forming units (BFU). Sufficient for performing at least 25 gel shift assays under standard conditions.
FORMULATION:	Liquid. In 20mM HEPES, pH 7.9, containing 25% glycerol, 420mM sodium chloride, 0.2mM EDTA and 1.5mM magnesium chloride.
SPECIFICITY:	Binds specifically to an oligonucleotide with the sequence GCG GGG GCG.
SPECIFIC ACTIVITY:	>0.33BFU/µl. 1BFU is sufficient to generate a band shift with a labelled oligonucleotide.
APPLICATION:	EMSA promoter characterization, in vitro transcription assay, analysis of nuclear extracts (oligonucleotides and recombinant protein serve as positive control).
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C
HANDLING:	Avoid freeze/thaw cycles.

Product Description

The recombinant EGR-3 protein can be employed in the following fields of research: a) Promoter studies: Identification of EGR-3 binding promoter elements. b) Detection of EGR-3 interacting proteins. c) Signal transduction: Assay for the effect on the nuclear component EGR-3. d) Screening for EGR-3 activity in tumor cells and lines. e) Drug screening: Effect on EGR-3 generation and DNA binding.

Product Specific Literature References

EGR3, a novel member of the Egr family of genes encoding immediate- early transcription factors: S. Patwardhan, et al.; Oncogene 6, 917 (1991) Abstract
Coordinate expression and distinct DNA-binding characteristics of the four EGR-zinc finger proteins in Jurkat T lymphocytes: C. Skerka, et al.; Immunobiology 198, 179 (1997) Abstract
The human zinc finger protein EGR-4 acts as autoregulatory transcriptional repressor: P.F. Zipfel, et al.; Biochim. Biophys. Acta 354, 134 (1997) Abstract
The early growth response protein (EGR-1) regulates interleukin-2 transcription by synergistic interaction with the nuclear factor of activated T cells: E.L. Decker, et al.; J. Biol. Chem. 273, 26923 (1998) Abstract; Full Text
Cyclosporin A-sensitive transcription factor Egr-3 regulates Fas ligand expression: P.R. Mittelstadt & J.D. Ashwell; Mol. Cell. Biol. 18, 3744 (1998) Abstract; Full Text
Role of Egr-2 in up-regulation of Fas ligand in normal T cells and aberrant double-negative lpr and gld T cells: P.R. Mittelstadt & J.D. Ashwell; J. Biol. Chem. 274, 3222 (1999) Abstract; Full Text

General Information

Early Growth Response- (EGR-) proteins represent a family of related transcription factors with almost identical DNA- binding zinc finger domains, but with different flanking regions. All four human EGR-proteins, EGR-1 to EGR-4 bind the EGR-consensus sequence GCG T/GGG GCG. However individual EGR-proteins show different binding to related sequences, such as the overlapping EGR-1/Sp1 binding site in the human IL-2 gene promoter. In functional assays the EGR-4 protein shows superior performance for Electrophoretic Mobility Shift Assay (EMSA) compared to proteins expressed in E.coli.

BACKGROUND/TECHNICAL INFORMATION

For EMSA labeled oligonucleotides are required, that include the specific target sequence. Radioactive labeling of the oligonucleotide is performed by kinase reaction (T4 kinase) using either 32P-γ ATP or 33P-γ ATP as a source for radioactivity. Due to the lower radiation load we recommend the use of 33P-γ ATP, which results in specific, detectable bands. Formation of a specific protein: DNA complex results in a lower mobility of this complex compared to the mobility of the free DNA. The specificity of the EMSA can also be analyzed by competition experiments in which an excess of unlabeled specific oligonucleotide interferes with the formation of a specific band, while the same amount of unrelated or mutated oligonucleotide has no effect. DNA-Protein complexes are then separated from the free (unbound) oligonucleotides by native gel-electrophoresis.

Swiss-Prot link Q06889: EGR-3 (human)

Further Categories Containing This Product:

Recombinant Proteins/Fusion Proteins

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