The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
EGR3 is a member of the early growth response transcription factor family of C2H2 zinc finger proteins (other members EGR1, EGR2 and EGR4). EGR proteins are immediate early proteins, expression of which is swiftly upregulated in response to a wide range of extracellular stimuli. EGR3 is also thought to be involved in development of muscle spindles, and is upregulated in several regions of the brain in response to stress or injury.
ICC/IF image of ab75461 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75461, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa cells at 5µg/ml.
Kearney SJ et al. Type I IFNs downregulate myeloid cell IFN-? receptor by inducing recruitment of an early growth response 3/NGFI-A binding protein 1 complex that silences ifngr1 transcription. J Immunol191:3384-92 (2013).
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