This antibody gave a positive signal in Human Heart Mitochondrial Lysate within Western Blot.
It also gave a positive signal in HepG2 cell line in IF and in FFPE human heart tissue sections in IHC.
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.
pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
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 before commencing with IHC staining protocol.
Use a concentration of 10 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 77 kDa (predicted molecular weight: 86 kDa).
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. MFN2 acts independently of the cytoskeleton. It therefore plays a central role in mitochondrial metabolism and may be associated with obesity and/or apoptosis processes. Overexpression induces the formation of mitochondrial networks. Plays an important role in the regulation of vascular smooth muscle cell proliferation. Involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). Is required for PARK2 recruitment to dysfunctional mitochondria. Involved in the control of unfolded protein response (UPR) upon ER stress including activation of apoptosis and autophagy during ER stress. Acts as an upstream regulator of EIF2AK3 and suppresses EIF2AK3 activation under basal conditions.
Ubiquitous; expressed at low level. Highly expressed in heart and kidney.
Involvement in disease
Charcot-Marie-Tooth disease 2A2 Neuropathy, hereditary motor and sensory, 6A
Belongs to the TRAFAC class dynamin-like GTPase superfamily. Dynamin/Fzo/YdjA family. Mitofusin subfamily. Contains 1 dynamin-type G (guanine nucleotide-binding) domain.
Phosphorylated by PINK1. Ubiquitinated by non-degradative ubiquitin by PARK2, promoting mitochondrial fusion; deubiquitination by USP30 inhibits mitochondrial fusion.
Mitochondrion outer membrane. Colocalizes with BAX during apoptosis.
Western blot - Anti-Mitofusin 2 antibody (ab101055)This image is courtesy of an anonymous abreview.
Anti-Mitofusin 2 antibody (ab101055) reactivity with reduced Rat primary neuron cell lysate after SDS-PAGE. Membranes were blocked in 5% milk in TBST+ 0.05% Tween for 1h at 25ºC before incubation with ab101055 (1μg/ml dilution in 5% BSA in PBS + 0.05% Tween) for 16h at 4ºC. The blot was then incubated with an anti-Rabbit HRP-conjugated secondary antibody before developing with ECL
ICC/IF image of ab101055 stained HepG2 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 (ab101055, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa and HepG2 cells at 10µg/ml, and in 100% methanol fixed (5 min) HeLa and MCF7 cells at 10µg/ml.
Western blot - Anti-Mitofusin 2 antibody (ab101055)
All lanes : Anti-Mitofusin 2 antibody (ab101055) at 1 µg/ml
Lane 1 : Human Heart Mitochondrial Lysate at 5 µg Lane 2 : Human Heart Mitochondrial Lysate at 10 µg
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
IHC image of ab101055 staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab101055, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer 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.
Peyta L et al. Reduced cardiolipin content decreases respiratory chain capacities and increases ATP synthesis yield in the human HepaRG cells. Biochim Biophys Acta1857:443-53 (2016).
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