The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 52 kDa).
1/50 - 1/100.
1/50 - 1/100.
May be a regulated effector of stress granule assembly. Phosphorylation-dependent sequence-specific endoribonuclease in vitro. Cleaves exclusively between cytosine and adenine and cleaves MYC mRNA preferentially at the 3'-UTR. ATP- and magnesium-dependent helicase. Unwinds preferentially partial DNA and RNA duplexes having a 17 bp annealed portion and either a hanging 3' tail or hanging tails at both 5'- and 3'-ends. Unwinds DNA/DNA, RNA/DNA, and RNA/RNA substrates with comparable efficiency. Acts unidirectionally by moving in the 5' to 3' direction along the bound single-stranded DNA.
Phosphorylated exclusively on serine residues. Hyperphosphorylated in quiescent fibroblasts. Hypophosphorylation leads to a decrease in endoribonuclease activity (By similarity). RASA1-dependent phosphorylation of Ser-149 induces a conformational change that prevents self-association. Dephosphorylation after HRAS activation is required for stress granule assembly. Ser-149 phosphorylation induces partial nuclear localization. Arg-435 is dimethylated, probably to asymmetric dimethylarginine.
Cytoplasm. Cytoplasm > cytosol. Cell membrane. Nucleus. Cytoplasmic in proliferating cells, can be recruited to the plasma membrane in exponentially growing cells (By similarity). Cytosolic and partially nuclear in resting cells. Recruited to stress granules (SGs) upon either arsenite or high temperature treatment. Recruitment to SGs is influenced by HRAS.
Immunohistochemical analysis of paraffin-embedded Human colon tissue staining G3BP using ab181150 at 1/100 dilution, and prediluted HRP Polymer for Rabbit IgG as a secondary antibody with Hematoxylin counterstain.
Immunofluorescent analysis of 4% paraformaldehyde fixed 293 cells staining G3BP using ab181150 at 1/100 dilution, and Alexa Fluor®555 stained Goat anti Rabbit IgG at 1/200 dilution as a secondary antibody (red). Dapi counterstain (blue)
Western blot analysis of G3BP in immunoprecipitation pellets from Ramos cell lysate, using ab181151 at a 1/50 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as a secondary antibody at 1/1000 dilution. Blocking/dilution buffer and concentration: 5% NFDM/TBST
Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/50 dilution + Immunoprecipitate from Ramos cell lysate using ab181150 with NFDM/TBST at 5 %
Secondary Peroxidase conjugated Goat anti-Rabbit IgG (H+L) at 1/1000 dilution