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 27 kDa (predicted molecular weight: 27 kDa).
FunctionBinds to the 60S ribosomal subunit and prevents its association with the 40S ribosomal subunit to form the 80S initiation complex in the cytoplasm. May behave as a stimulatory translation initiation factor downstream insulin/growth factors. Is also involved in ribosome biogenesis. Associates with pre-60S subunits in the nucleus and is involved in its nuclear export. Cytoplasmic release of TIF6 from 60S subunits and nuclear relocalization is promoted by a RACK1 (GNB2L1)-dependent protein kinase C activity.
Tissue specificityExpressed at very high levels in colon carcinoma with lower levels in normal colon and ileum and lowest levels in kidney and muscle (at protein level).
Sequence similaritiesBelongs to the eIF-6 family.
Post-translational modificationsPhosphorylation at Ser-174 and Ser-175 promotes nuclear export.
Cellular localizationCytoplasm. Nucleus > nucleolus. Shuttles between cytoplasm and nucleus/nucleolus.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 27 kDa Observed band size : 27 kDa Additional bands at : 95 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 90 seconds
Immunocytochemistry/ Immunofluorescence - integrin beta 4 binding protein antibody (ab77298)
ICC/IF image of ab77298 stained Hek293 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 (ab77298, 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) HeLa, HepG2, and MCF-7 cells at 5µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, and MCF-7 cells at 1µg/ml.
References for Anti-integrin beta 4 binding protein antibody (ab77298)
has not yet been referenced specifically in any publications.
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