WB
Western blot analysis of MFF on different lysates with Rabbit anti-MFF antibody at 1/25,000 dilution. Lane 1: HeLa cell lysate (10 µg/Lane) Lane 2: MCF7 cell lysate (10 µg/Lane) Lane 3: A549 cell lysate (10 µg/Lane) Lane 4: HepG2 cell lysate (10 µg/Lane) Lane 5: 293T cell lysate (10 µg/Lane) Lane 6: C2C12 cell lysate (10 µg/Lane) Lane 7: PC-12 cell lysate (10 µg/Lane) Lane 8: COS-1 cell lysate (10 µg/Lane) Lane 9: Mouse brain tissue lysate (20 µg/Lane) Lane 10: Mouse heart tissue lysate (20 µg/Lane) Lane 11: Rat brain tissue lysate (20 µg/Lane) Lane 12: Rat heart tissue lysate (20 µg/Lane) Predicted band size: 38 kDa Observed band size: 30-38 kDa Exposure time: 4 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/25,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-MFF antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling MFF with Rabbit anti-MFF antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MFF antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells labeling MFF. Cells were fixed and permeabilized. Then stained with the primary antibody (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
MFF was immunoprecipitated from 0.2 mg HeLa cell lysate with at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: IP in HeLa cell lysate Lane 3: Rabbit IgG instead of in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 7 seconds; ECL: K1801| Product Name | MFF Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human MFF aa 1-322. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCIFIHCIPWB |
| WB:1:25,000 IHC:1:5,000 FC:1:1,000 IF:1:100 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | MFF |
|---|---|
| Gene Synonyms | EMPF2 GL004 C2orf33 |
| Gene Full Name | mitochondrial fission factor |
| Gene Summary | This is a nuclear gene encoding a protein that functions in mitochondrial and peroxisomal fission. The encoded protein recruits dynamin-1-like protein (DNM1L) to mitochondria. There are multiple pseudogenes for this gene on chromosomes 1, 5, and X. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Mar 2013] |
| Alternative Names | C2orf33 antibody Chromosome 2 open reading frame 33 antibody DKFZp666J168 antibody GL004 antibody Mff antibody MFF_HUMAN antibody MGC110913 antibody Mitochondrial fission factor antibody OTTHUMP00000164235 antibody |
| Molecular Weight(MW) | 38 kDa |
| Cellular Localization | Cytoplasmic vesicle,Membrane,Mitochondrion,Mitochondrion outer membrane,Peroxisome,Synapse. |
WB
Western blot analysis of MFF on different lysates with Rabbit anti-MFF antibody at 1/25,000 dilution. Lane 1: HeLa cell lysate (10 µg/Lane) Lane 2: MCF7 cell lysate (10 µg/Lane) Lane 3: A549 cell lysate (10 µg/Lane) Lane 4: HepG2 cell lysate (10 µg/Lane) Lane 5: 293T cell lysate (10 µg/Lane) Lane 6: C2C12 cell lysate (10 µg/Lane) Lane 7: PC-12 cell lysate (10 µg/Lane) Lane 8: COS-1 cell lysate (10 µg/Lane) Lane 9: Mouse brain tissue lysate (20 µg/Lane) Lane 10: Mouse heart tissue lysate (20 µg/Lane) Lane 11: Rat brain tissue lysate (20 µg/Lane) Lane 12: Rat heart tissue lysate (20 µg/Lane) Predicted band size: 38 kDa Observed band size: 30-38 kDa Exposure time: 4 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/25,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-MFF antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling MFF with Rabbit anti-MFF antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MFF antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling MFF. Cells were fixed and permeabilized. Then stained with the primary antibody (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
MFF was immunoprecipitated from 0.2 mg HeLa cell lysate with at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: IP in HeLa cell lysate Lane 3: Rabbit IgG instead of in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 7 seconds; ECL: K1801| Application Notes | WB:1:25,000 IHC:1:5,000 FC:1:1,000 IF:1:100 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
