ALEXIS Biochemicals: Sp1 Protein (human) (recombinant) (HA)

ALX-201-106

Revised 08-Mar-07

Sp1 Protein (human) (recombinant) (HA)
PRODUCT LINE	DNA Regulation / Transcription
PRODUCT CATEGORY	Sp Protein Family / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-106-R050		50 µl	377.00 USD

Product Specification

SOURCE/HOST:	Produced in insect cells (Schneider SL2 cells). Human Sp1 is fused to a HA- and a FLAG^®-tag (H. Braun and G. Suske; Biotechniques 26, 1038 (1999)).
QUANTITY:	Sufficient for performing at least 25 protein-DNA interaction experiments in 10mM HEPES, pH 7.9, 0.1mM EDTA, 8.5% glycerol, 170mM KCl, 1µM ZnSO₄, 37.5µg/ml Poly dIdC, 10mM DTT and 1mg/ml BSA, analyzed by gel shift assay.
PURITY DETAIL:	DNA-affinity purified.
FORMULATION:	Liquid. In 50mM TRIS-HCl, pH 7.9, containing 1M KCl, 0.1% NP-40, 12.5mM MgCl₂and 20% glycerol.
SPECIFIC ACTIVITY:	0.5BFU/µl (band forming units). 1BFU is sufficient to generate a strong band shift with a labelled oligonucleotide.
APPLICATION:	EMSA promoter characterization, in vitro transcription assays, analysis of nuclear extracts (oligonucleotides and recombinant protein serve as a positive control).
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C
HANDLING:	Avoid freeze/thaw cycles.

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Product Description

The ubiquitously expressed Sp1 was the first identified and cloned member of the Sp-family of mammalian transcription factors. The 778 aa human protein contains an 81 aa DNA-binding domain consisting of three C₂H₂-type zinc fingers close to the C-terminus. Two glutamine-rich activation domains are present adjacent to serine/threonine stretches in the N-terminal part of the protein. Sp1 recognizes specifically G-rich elements such as the GC-box (GGGGCGGGG) and the GT/CACC-box (GGTGTGGGG) found in housekeeping as well as in many tissue-specific and viral genes. The protein may be employed in the following fields of research:
i) Promoter studies: Identification of Sp1 binding promoter elements by bandshift or DNase1
footprint assays.
ii) Promoter studies: Activation by Sp1 in in vitro transcription assays.
iii) Detection of Sp1 interacting proteins.

Product Specific Literature References

Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain: J.T. Kadanoga, et al.; Cell 51, 1079 (1987) Abstract
Vectors for inducible expression of dual epitope-tagged proteins in insect cells: H. Braun & G. Suske; Biotechniques 26, 1038 (1999) Abstract
The Sp-family of transcription factors: G. Suske; Gene 238, 291 (1999), (Review) Abstract

General Information

BACKGROUND/TECHNICAL INFORMATION

Nuclear extract was prepared from HA/FLAG-Sp1 expressing SL2 cells and precleared by unspecific DNA-affinity precipitation (multimerized oligonucleotide with HNF-3 binding site). Sp1 was then precipitated with a specific DNA-affinity matrix (multimerized double-stranded GC-box oligonucleotide with the sequence 5´-AGCTTCCGTTGGGGCGGGGCTTCACGTCGA-3´). Bound protein was eluted with 50mM TRIS-HCl, pH 7.9, containing 1M KCl, 0.1% NP40, 20% glycerol, 12.5mM MgCl₂.
In EMSAs, DNA-protein complexes are separated from free (unbound) oligonucleotides by native gel-electrophoresis. The formation of a specific DNA-protein complex results in a lower mobility of this complex compared to the mobility of the free DNA.
The specificity of the DNA-protein interaction can be analyzed by competition or supershift experiments. An excess of unlabeled specific oligonucleotide interferes with the formation of a specific band, whereas unspecific or mutated oligonucleotides have no effect. The presence of a specific antibody leads to the formation of a very slow migrating antibody-protein-DNA complex.

FLAG is a registered trademark of Sigma-Aldrich Co.

General Literature References

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Further Categories Containing This Product:

Recombinant Proteins / Fusion Proteins

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