Product nameTiam1 Activation Assay Kit
Sample typeTissue, Adherent cells, Suspension cells
Species reactivityReacts with: Human
Tiam 1 Activation Assay Kit (ab211173) uses nucleotide-free Rac1 G15A agarose beads to selectively isolate and pull down the active form of Rac-GEF from cell or tissue lysates. The precipitated active Rac-GEF is subsequently detected by immunoblot analysis using an anti Tiam-1 antibody, which reacts with the human protein. Tiam-1 is a specific GEF known to activate Rac1.
Features: 1) non radioactive assay format; 2) fast results: 1 hour assay plus electrophoresis/blotting time; 3) includes Cdc42 positive control; 4) pink colored agarose beads for easy identification during washing and aspiration steps.
Small GTP-binding proteins (or GTPases) are a family of proteins that serve as molecular regulators in signaling transduction pathways.
Rac, a 21 kDa protein, belongs to the family of Rho GTPases and is involved in regulating a variety of biological pathways that such as cell motility, cell division, gene transcription, and cell transformation. Like other small GTPases, Rac influences molecular events by cycling between an inactive GDP-bound form and an active GTP-bound form.
The cycling process between GDP-bound and GTP-bound state is regulated primarily by two distinct families of proteins: guanine nucleotide exchange factors (GEFs), which activate Rho proteins by catalyzing the exchange of GDP for GTP, and the GTPase activating proteins or GAPs, which negatively regulate GTPase function by stimulating GTP hydrolysis.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 20 tests 5X Assay/Lysis Buffer 1 x 30ml Anti-Tiam1 Rabbit polyclonal antibody 1 x 40µl Rac1 G15A Agarose Beads (Active Rac-GEF) 1 x 800µl
FunctionModulates the activity of RHO-like proteins and connects extracellular signals to cytoskeletal activities. Acts as a GDP-dissociation stimulator protein that stimulates the GDP-GTP exchange activity of RHO-like GTPases and activates them. Activates RAC1, CDC42, and to a lesser extent RHOA.
Tissue specificityFound in virtually all analyzed tumor cell lines including B- and T-lymphomas, neuroblastomas, melanomas and carcinomas.
Sequence similaritiesBelongs to the TIAM family.
Contains 1 DH (DBL-homology) domain.
Contains 1 PDZ (DHR) domain.
Contains 2 PH domains.
Contains 1 RBD (Ras-binding) domain.
Cellular localizationCell junction. Presence of KRIT1, CDH5 and RAP1B is required for its localization to the cell junction.
- Information by UniProt
- Human T-lymphoma invasion and metastasis inducing TIAM1 protein
- T cell lymphoma invasion and metastasis 1
- T lymphoma invasion and metastasis inducing TIAM1 protein
Tiam1 (Rac-GEF) activation assay in MDA-231 Cells. Active Tiam-1 was pulled down from MDA- 231 lysate (2 mg) with Rac1 G15A agarose beads and probed with anti-Tiam1 antibody.
Tiam1 activation assay in 293 cells. 293 cells were transfected with active Tiam1 (ΔN Tiam1). Active Tiam1 in lysate was pulled down with Rac1 G15A agarose beads. Lane 1, Mock Transfection Control. Lane 2, ΔN Tiam1 Transfection.
ab211173 has not yet been referenced specifically in any publications.