The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50 - 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Use buffer at pH 6 for antigen retrieval.
Use a concentration of 1 - 4 µg/ml. Fixation/Permeabilization: PFA/Triton X-100
1/250 - 1/500.
FunctionFunctions as a calcium-regulated nucleotide exchange factor activating both Ras and RAC1 through the exchange of bound GDP for GTP. Preferentially activates HRAS in vivo compared to RRAS based on their different types of prenylation. Functions in synaptic plasticity by contributing to the induction of long term potentiation.
Tissue specificityWidely expressed with higher expression in brain, followed by heart, lung, pancreas and kidney. Detected in placenta. Expressed in brain and lung (at protein level).
DomainThe Ras-GEF domain and the N-terminal Ras-GEF domain form a Ras-binding site and mediate Ras activation. The IQ domain mediates the calcium-dependent interaction with calmodulin but is dispensable for the Ras-GEF activity. The DH (DBL-homology) domain mediates interaction with RASGRF1 and EPB49 and is required for RAC1 activation.
Post-translational modificationsPhosphorylated by CDK5; down-regulates RASGRF2-mediated RAC1 activation. Ubiquitinated upon interaction with Ras. Ubiquitination leads to degradation through the 26S proteasome.
Cellular localizationCytoplasm. Cell membrane. Endoplasmic reticulum membrane. Translocates to membranes when activated. Found both at cell periphery and along the axon of neurons.