The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use at a concentration of 5 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Actin-binding adapter protein. May act as a common effector of antigen receptor-signaling pathways in leukocytes. Its association with dynamin suggests that it may also connect the actin cytoskeleton to endocytic function. Acts as a key component of the immunological synapse that regulates T-cell activation by bridging TCRs and the actin cytoskeleton to gene activation and endocytic processes. Binds to F-actin but is not involved in actin polymerization, capping or bundling. Does not bind G-actin.
Belongs to the ABP1 family. Contains 1 ADF-H domain. Contains 1 SH3 domain.
The SH3 domain mediates interaction with SHANK2, SHANK3 and PRAM1.
src homology 3 domain containing protein HIP 55 antibody
Western blot - HIP55 antibody (ab77147)
All lanes : Anti-HIP55 antibody (ab77147) at 1 µg/ml
Lane 1 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate Lane 2 : SK N BE (Human neuroblastoma) Whole Cell Lysate Lane 3 : SK N SH (Human neuroblastoma) Whole Cell Lysate Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
ICC/IF image of ab77147 stained HeLa 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 (ab77147, 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, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.