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Sp Protein Family / Related Products
You are here: Product Lines > DNA Regulation / Transcription > Transcription Factors / Related Products > Sp Protein Family / Related Products
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ALX-201-106 Revised 08-Mar-07
Sp1 Protein (human) (recombinant) (HA)
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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 Add To Cart
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 MgCl2 and 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
For an extended bibliography click here.:
Further Categories Containing This Product:
Recombinant Proteins / Fusion Proteins
 
 
ALX-201-107 Revised 17-Nov-06
Sp3 Protein (human) (recombinant) (HA)
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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-201-107-R050   50 µl 377.00 USD Add To Cart
Product Specification
SOURCE/HOST: Produced in insect cells (Schneider SL2 cells). Human Sp3 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 10 mM HEPES (pH 7.9), 0.1 mM EDTA, 8.5 % Glycerol, 170 mM KCl, 1 µM ZnSO4, 37.5 µg/ml Poly dIdC, 10 mM DTT, and 1 mg/ml BSA, analyzed by gel shift assay.
PURITY: ~50% (SDS-PAGE)
FORMULATION: Liquid. In 20mM TRIS-Cl, pH 7.9, containing 150mM KCl, 0.2mM EDTA and 20% glycerol.
SPECIFICITY: Binds specifically to an oligonucleotide with the sequence: GGG GCG GGG.
SPECIFIC ACTIVITY: >0.5 BFU/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
Product Specific Literature References
The Sp-family of transcription factors: G. Suske; Gene 238, 291 (1999), (Review) Abstract
Vectors for inducible expression of dual epitope-tagged proteins in insect cells: H. Braun & G. Suske; Biotechniques 26, 1038 (1999) Abstract
General Information
BACKGROUND/TECHNICAL INFORMATION FLAG is a registered trademark of Sigma-Aldrich Co.
Further Categories Containing This Product:
Recombinant Proteins / Fusion Proteins
 
 
ALX-380-064 Revised 08-Oct-08
WP631 . dihydrochloride
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PRODUCT LINE Natural Products / Antibiotics
PRODUCT CATEGORY Other Antibiotics - DNA Regulation / Transcription
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-380-064-C100   100 µg 60.00 USD Add To Cart
ALX-380-064-M001   1 mg 240.00 USD Add To Cart
Product Specification
FORMULA: C62H64O20N2 . 2HCl
MW: 1157.2 . 73.0
PURITY: ≥95%
APPEARANCE: Off-white to red powder.
SOLUBILITY: Soluble in DMSO or dimethyl formamide.
SHIPPING: AMBIENT
LONG TERM STORAGE: +4°C
USE/STABILITY: Dissolve the compound in DMSO or dimethyl formamide at 20mg/ml, then add water (or 5% dextrose, PBS, etc.) to bring it to a final concentration of 1mg/ml (5% DMSO or dimethyl formamide).
HANDLING: Keep cool and dry.
HAZARD: HARMFUL. MAY BE CARCINOGENIC.

Product Description
Synthetic fluorescent bisintercalating anthracycline antibiotic with ultratight binding properties to DNA. Inhibits Sp1-initiated transcription at nanomolar concentrations. Appears to overcome MRP-mediated multidrug resistance. Induces apoptosis. Licenced product covered by US Patent 5,874,412. Sold with permission of Houston Pharmaceuticals, Inc.
Product Specific Literature References
Structure-based design of a new bisintercalating anthracycline antibiotic: J.B. Chaires, et al.; J. Med. Chem. 40, 261 (1997) Abstract
Structure of a DNA-bisdaunomycin complex: G.G. Hu, et al.; Biochemistry 36, 5940 (1997) Abstract
Bisanthracycline WP631 inhibits basal and Sp1-activated transcription initiation in vitro: B. Martin, et al.; Nucl. Acids Res. 27, 3402 (1999) Abstract; Full Text
Analysis of the effects of Daunorubicin and WP631 on transcription: J. Portugal, et al.; Curr. Med. Chem. 8, 1 (2001), (Review) Abstract
Identification of a critical Sp1 site within the endoglin promoter and its involvement in the transforming growth factor-beta stimulation: L.M. Botella, et al.; J. Biol. Chem. 276, 34486 (2001) Abstract; Full Text
Exploiting anthracycline scaffold for designing DNA-targeting agents: W. Priebe; Methods Enzymol. 340, 529 (2001) Abstract
Induction of G(2)/M arrest and inhibition of c-myc and p53 transcription by WP631 in Jurkat T lymphocytes: S. Villamarin, et al.; Biochem. Pharmacol. 63, 1251 (2002) Abstract
Evidence that activation of nuclear factor-kappaB is essential for the cytotoxic effects of doxorubicin and its analogues: K. Ashikawa, et al.; Biochem. Pharmacol. 67, 353 (2004) Abstract
Bis-anthracycline antibiotics inhibit human immunodeficiency virus type 1 transcription: O. Kutsch, et al.; Antimicrob. Agents. Chemother. 48, 1652 (2004) Abstract
Circumvention of the multidrug-resistance protein (MRP-1) by an antitumor drug through specific inhibition of gene transcription in breast tumor cells: S. Mansilla, et al.; Biochem. Pharmacol. 73, 934 (2007) Abstract
Further Categories Containing This Product:
MDR Inhibitors • Antibiotics - Apoptosis Inducers & Inhibitors • Sp Protein Family / Related Products • DNA Intercalators & Crosslinkers
 
 
ALX-380-074 Revised 12-Oct-07
WP631 . dimethanesulfonate
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PRODUCT LINE Natural Products / Antibiotics
PRODUCT CATEGORY Other Antibiotics - DNA Regulation / Transcription
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-380-074-C500   500 µg 150.00 USD Add To Cart
Product Specification
FORMULA: C62H64N2O20 . 2CH3SO3
MW: 1157.2 . 191.2
PURITY: ≥95%
APPEARANCE: Red powder.
SOLUBILITY: Soluble in DMSO (10mg/ml) or water (1mg/ml).
SHIPPING: AMBIENT
LONG TERM STORAGE: +4°C
HAZARD: HARMFUL. MAY BE CARCINOGENIC.

Product Description
Synthetic fluorescent bisintercalating anthracycline antibiotic with ultratight binding properties to DNA. Inhibits Sp1-initiated transcription at nanomolar concentrations. Appears to overcome multidrug resistance protein (MRP) mediated multidrug resistance (MDR). More water soluble than WP631 . dihydrochloride (Prod. No. ALX-380-064). Licenced product covered by US Patent 5,874,412. Sold with permission of Houston Pharmaceuticals, Inc.
Product Specific Literature References
Structure-based design of a new bisintercalating anthracycline antibiotic: J.B. Chaires, et al.; J. Med. Chem. 40, 261 (1997) Abstract
Structure of a DNA-bisdaunomycin complex: G.G. Hu, et al.; Biochemistry 36, 5940 (1997) Abstract
Binding of two novel bisdaunorubicins to DNA studied by NMR spectroscopy: H. Robinson, et al.; Biochemistry 36, 8663 (1997) Abstract
Ultratight DNA binding of a new bisintercalating anthracycline antibiotic: F. Leng, et al.; Biochemistry 37, 1743 (1998) Abstract
Drug-DNA interactions: J.B. Chaires; Curr. Opin. Struct. Biol. 8, 314 (1998) Abstract
Bisanthracycline WP631 inhibits basal and Sp1-activated transcription initiation in vitro: B. Martin, et al.; Nucleic Acids Res. 27, 3402 (1999) Abstract
Analysis of the effects of daunorubicin and WP631 on transcription: J. Portugal, et al.; Curr. Med. Chem. 8, 1 (2001) Abstract
Inhibition of basal and transforming growth factor-beta-induced stimulation of COL1A1 transcription by the DNA intercalators, mitoxantrone and WP631, in cultured human dermal fibroblasts: S. Gaidarova and S.A. Jimenez; J. Biol. Chem. 277, 38737 (2002) Abstract
Sequence selective binding of bis-daunorubicin WP631 to DNA: K.R. Fox, et al.; Eur. J. Biochem. 271, 3556 (2004) Abstract
Further Categories Containing This Product:
DNA Intercalators & Crosslinkers • MDR Inhibitors • Sp Protein Family / Related Products • Antibiotics - Apoptosis Inducers & Inhibitors
 
 
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