The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent dilution. PubMed: 20008164
1/64000. Peptide ELISA
Use a concentration of 0.003 - 0.01 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 61 kDa).
Plays a role in membrane reorganization in response to nucleotide hydrolysis. Binds to liposomes and deforms them into tubules. Plays a role in membrane trafficking between the plasma membrane and endosomes. Important for the internalization of GLUT4. Required for normal fusion of myoblasts to skeletal muscle myotubes. Binds ATP; does not bind GTP.
Highly expressed in heart and moderately expressed in placenta, lung, and skeletal muscle.
Contains 1 EF-hand domain. Contains 1 EH domain.
The EH domain interacts with Asn-Pro-Phe (NPF) motifs of target proteins.
Cell membrane. Endosome membrane. Colocalizes with GLUT4 in intracellular tubulovesicular structures that are associated with cortical F-actin.
ICC/IF image of ab23935 stained A549 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 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 (ab23935, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96931, DyLight® 488 donkey anti-goat IgG (H+L) used at a 1/250 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
Western blot - EHD2 antibody (ab23935)
Anti-EHD2 antibody (ab23935) at 0.003 µg/ml + Human Heart lysate (RIPA buffer, 35µg total protein per lane).
Predicted band size : 61 kDa Observed band size : 61 kDa Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
Western blot - Anti-EHD2 antibody (ab23935)
Anti-EHD2 antibody (ab23935) at 0.1 µg/ml + Human Placenta lysate in RIPA buffer at 35 µg