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Active Recombinant Human TIAM1 Protein

Cat.No. : TIAM1-25H
Product Overview : The DH/PH domain of human Tiam1 protein has been produced in a bacterial expression system. It contains an MBP fusion at its amino terminus.
  • Specification
  • Gene Information
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Source : E. coli
Species : Human
Form : White lyophilized powder.
Bio-activity : The biological activity of Tiam1 DHPH can be determined from its ability to catalyze nucleotide exchange on Rac1 using the nucleotide exchange assay of Bodipy-GDP for excess GDP or GTP. Rac1 protein is pre-loaded with Bodipy-FL-GDP by adding excess EDTA e.g. 0.7 mmol EDTA per mmol Mg2+ ions present in the reaction. This sub-stock solution is then used in a dissociation assay format which indicates competition for exchange site with unlabeled nucleotide. The reaction is monitored by fluorescence measurement at 485nm Ex / 535nm Em. Stringent quality control ensures that the exchange rate of Bodipy-GDP or mant-GDP is enhanced at least five fold in the presence of 0.8 µM Tiam1 DHPH.
Molecular Mass : 86 kDa
Purity : Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% polyacrylamide gradient gel. MBP-Tiam1 was determined to be 90% pure.
Storage : The protein is stable for six months if stored at -70 centigrade.
Storage Buffer : 1. MBP-Tiam1 DH/PH protein.
2. Rac1 protein.
3. Exchange buffer 2 (20 mM Tris-HCl pH 7.5, 50 mM NaCl, 1 mM DTT, 2 mM EDTA, 100 µg/ml BSA, and 0.75 µM Bodipy-FL-GDP), note - make fresh.
4. 50 mM MgCl2 in 20 mM Tris-HCl pH 7.5, 50 mM NaCl.
5. 5 mM GTP in 20 mM Tris-HCl pH 7.5, 50 mM NaCl.
Reconstitution : Before reconstitution, briefly centrifuge to collect the product at the bottom of the tube. The protein should be reconstituted to 5 mg/ml with ice cold nanopure water (20 µl water per 100 µg protein). When reconstituted, the protein will be in the following buffer: 20 mM Tris pH 7.5, 50 mM NaCl, 1 mM MgCl2, 5% (v/v) sucrose and 1% (v/v) dextran. In order to maintain high biological activity of the protein it is strongly recommended that the protein solution be supplemented with DTT to 1 mM final concentration, aliquoted into "experiment sized" amounts, snap frozen in liquid nitrogen and stored at -70centigrade. The protein must not be exposed to repeated freeze-thaw cycles. The lyophilized protein is stable at 4centigrade desiccated (<10% humidity) for one year. Further Tiam1 dilutions should be made in 20 mM Tris pH 7.5, 50 mM NaCl, 1 mM MgCl2 (not supplied).
Gene Name : TIAM1
Official Symbol : TIAM1 T cell lymphoma invasion and metastasis 1 [ Homo sapiens (human) ]
Synonyms : TIAM1; T cell lymphoma invasion and metastasis 1; TIAM-1;
Gene ID : 7074
mRNA Refseq : NM_003253
Protein Refseq : NP_003244
MIM : 600687
UniProt ID : Q13009

For Research Use Only. Not intended for any clinical use. No products from Creative BioMart may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative BioMart.

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