WB
Western blot analysis of OPA1 on different lysates with Rabbit anti-OPA1 antibody at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: HeLa treated with 10μM CCCP for 30 minutes cell lysate (20 µg/Lane), Lane 3: MCF7 cell lysate (20 µg/Lane), Lane 4: LNCaP cell lysate (20 µg/Lane), Lane 5: PANC-1 cell lysate (20 µg/Lane), Lane 6: 293T cell lysate (20 µg/Lane), Lane 7: NIH/3T3 cell lysate (20 µg/Lane), Lane 8: C2C12 cell lysate (20 µg/Lane), Lane 9: Neuro-2a cell lysate (20 µg/Lane), Lane 10: PC-12 cell lysate (20 µg/Lane), Lane 11: C6 cell lysate (20 µg/Lane), Lane 12: Mouse brain tissue lysate (40 µg/Lane), Lane 13: Mouse liver tissue lysate (40 µg/Lane), Lane 14: Rat brain tissue lysate (40 µg/Lane), Exposure time: 2 seconds; 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 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST 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 kidney tissue with Rabbit anti-OPA1 antibody at 1/1,000 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 1/1,000 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.IHC
Immunohistochemical analysis of paraffin-embedded rat retina tissue with Rabbit anti-OPA1 antibody at 1/1,000 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 1/1,000 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 OPA1 with Rabbit anti-OPA1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-OPA1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells labeling OPA1. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (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
OPA1 was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-OPA1 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-OPA1 antibody at 1/1,000 dilution. 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: Rabbit anti-OPA1 antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-OPA1 antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 10 seconds.| Product Name | OPA1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description |
|
| Immunogen | Recombinant protein within human OPA1 aa 201-960. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:1000-1:5000 IHC:1:1000 ICC/IF:1:100 FC:1:1000 IP: 1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | OPA1 |
|---|---|
| Gene Synonyms | NPG NTG MGM1 BERHS largeG MTDPS14 |
| Gene Full Name | OPA1 mitochondrial dynamin like GTPase |
| Gene Summary | The protein encoded by this gene is a nuclear-encoded mitochondrial protein with similarity to dynamin-related GTPases. The encoded protein localizes to the inner mitochondrial membrane and helps regulate mitochondrial stability and energy output. This protein also sequesters cytochrome c. Mutations in this gene have been associated with optic atrophy type 1, which is a dominantly inherited optic neuropathy resulting in progressive loss of visual acuity, leading in many cases to legal blindness. [provided by RefSeq, Aug 2017] |
| Molecular Weight(MW) | 112kDa(Observed band size: 80/100kDa) |
| Cellular Localization | Mitochondrion inner membrane. |
WB
Western blot analysis of OPA1 on different lysates with Rabbit anti-OPA1 antibody at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: HeLa treated with 10μM CCCP for 30 minutes cell lysate (20 µg/Lane), Lane 3: MCF7 cell lysate (20 µg/Lane), Lane 4: LNCaP cell lysate (20 µg/Lane), Lane 5: PANC-1 cell lysate (20 µg/Lane), Lane 6: 293T cell lysate (20 µg/Lane), Lane 7: NIH/3T3 cell lysate (20 µg/Lane), Lane 8: C2C12 cell lysate (20 µg/Lane), Lane 9: Neuro-2a cell lysate (20 µg/Lane), Lane 10: PC-12 cell lysate (20 µg/Lane), Lane 11: C6 cell lysate (20 µg/Lane), Lane 12: Mouse brain tissue lysate (40 µg/Lane), Lane 13: Mouse liver tissue lysate (40 µg/Lane), Lane 14: Rat brain tissue lysate (40 µg/Lane), Exposure time: 2 seconds; 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 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST 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 kidney tissue with Rabbit anti-OPA1 antibody at 1/1,000 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 1/1,000 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat retina tissue with Rabbit anti-OPA1 antibody at 1/1,000 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 1/1,000 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 OPA1 with Rabbit anti-OPA1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-OPA1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling OPA1. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (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
OPA1 was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-OPA1 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-OPA1 antibody at 1/1,000 dilution. 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: Rabbit anti-OPA1 antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-OPA1 antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 10 seconds.| Application Notes | WB:1:1000-1:5000 IHC:1:1000 ICC/IF:1:100 FC:1:1000 IP: 1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
