Recent lab testing showed that the antibody detects the band of interest at the proper molecular weight in several cell and tissue lysates. However, the antibody also detects a band at about 50kDa. We could not find in the literature any references on what this band could be. On our test, the 50kDa band detections decreased by decreasing the antibody concentration.
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Is unsuitable for IP.
Dynamin-related GTPase required for mitochondrial fusion and regulation of apoptosis. May form a diffusion barrier for proteins stored in mitochondrial cristae. Proteolytic processing in response to intrinsic apoptotic signals may lead to disassembly of OPA1 oligomers and release of the caspase activator cytochrome C (CYCS) into the mitochondrial intermembrane space.
Highly expressed in retina. Also expressed in brain, testis, heart and skeletal muscle. Isoform 1 expressed in retina, skeletal muscle, heart, lung, ovary, colon, thyroid gland, leukocytes and fetal brain. Isoform 2 expressed in colon, liver, kidney, thyroid gland and leukocytes. Low levels of all isoforms expressed in a variety of tissues.
Involvement in disease
Defects in OPA1 are a cause of optic atrophy type 1 (OPA1) [MIM:165500]. OPA1 is a dominantly inherited optic neuropathy occurring in 1 in 50,000 individuals that features progressive loss in visual acuity leading, in many cases, to legal blindness. Defects in OPA1 are the cause of optic atrophy 1 with deafness (OPA1D) [MIM:125250]. Some individuals with mutations in OPA1 manifest also ophthalmoplegia and myopathy.
Belongs to the dynamin family.
PARL-dependent proteolytic processing releases an antiapoptotic soluble form not required for mitochondrial fusion.
Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) labelling OPA1 with purified ab157457 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
ICC/IF image of ab157457 stained HCT116 cells. The cells were 100% methanol fixed (5 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 ab157457 at overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) 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 - Anti-OPA1 antibody [EPR11057(B)] (ab157457)
All lanes : Anti-OPA1 antibody [EPR11057(B)] (ab157457) at 1/1000 dilution
Lane 1 : HeLa cell lysate Lane 2 : HepG2 cell lysate Lane 3 : A431 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat anti-rabbit HRP at 1/2000 dilution