This antibody gave a positive signal in HeLa whole cell lysate and in the following tissue lysates: Human Kidney; Mouse Kidney; Rat Kidney; Human Heart; Mouse Heart; Rat Heart.
This antibody gave a positive result in IHC in the following FFPE tissue: Human Colon Adenocarcinoma.
This antibody gave a positive result when used in the following formaldehyde fixed cell lines: DU145.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Detects a band of approximately 84 kDa (predicted molecular weight: 84 kDa).
Use a concentration of 10 µg/ml.
Essential transmembrane GTPase, which mediates mitochondrial fusion. Fusion of mitochondria occurs in many cell types and constitutes an important step in mitochondria morphology, which is balanced between fusion and fission. MFN1 acts independently of the cytoskeleton. Overexpression induces the formation of mitochondrial networks.
Ubiquitous. Expressed at slightly higher level in kidney and heart. Isoform 2 may be overexpressed in some tumors, such as lung cancers.
ICC/IF image of ab107129 stained DU145 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 ab107129 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- chicken (ab96947) IgY (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.
IHC image of Mitofusin 1 staining in Human colon adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab107129, 5µg/ml, for 15 mins at room temperature. A Goat anti-Chicken biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-Mitofusin 1 antibody (ab107129)
All lanes : Anti-Mitofusin 1 antibody (ab107129) at 1 µg/ml
Lane 1 : Human kidney tissue lysate - total protein (ab30203) Lane 2 : Kidney (Mouse) Tissue Lysate Lane 3 : Kidney (Rat) Tissue Lysate Lane 4 : Human heart tissue lysate - total protein (ab29431) Lane 5 : Heart (Mouse) Tissue Lysate Lane 6 : Heart (Rat) Tissue Lysate Lane 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal Secondary Antibody to Chicken IgY - H&L (HRP) at 1/3000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 84 kDa Observed band size : 84 kDa Additional bands at : 17 kDa,27 kDa,36 kDa. We are unsure as to the identity of these extra bands.