The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent dilution. Detects a band of approximately 75 kDa (predicted molecular weight: 92 kDa). 1/1000 when using colorimetric substrates such as BCIP/NBT - 1/5000 for chemiluminescent substrates. EDTA/EGTA treatment of tissues or lysates is required to see latent zymogen.
Use a concentration of 1 µg/ml.
ADAM23 is a non-catalytic metalloprotease-like protein. It is highly expressed in the brain, primarily in the amygdala, caudate nucleus, hypothalamus, thalamus, cerebral cortex and occipital pole, and weakly expressed in the heart. 3 isoforms are produced by alternative splicing, alpha, beta and gamma. ADAM23 may play a role in cell-cell and cell-matrix interactions.
Single pass type I membrane protein.
3 isoforms produced by alternative splicing, gamma isoform is secreted.
ICC/IF image of ab28302 stained PC12 cells. The cells were 4% formaldehyde 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 (ab28302, 1µg/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.
Western blot - Anti-ADAM23 antibody - Cytoplasmic domain (ab28302)
All lanes : Anti-ADAM23 antibody - Cytoplasmic domain (ab28302) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466) Lane 2 : Rat Cortex Tissue Lysate Lane 3 : Human brain amygdala tissue lysate (alzheimer) - membrane extract (ab30118)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution