Phospho-DRP1 (S616) Recombinant Rabbit Monoclonal Antibody [PSH06-77]
cat.: HA722760
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Tissue, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH06-77 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 82 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser616 of human DRP1. |
| Positive control: | HeLa cell lysate, HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 100ng/mL nocodazole for 18 hours cell lysate, HeLa, C2C12 cell lysate, PC-12 cell lysate, mouse brain tissue, Jurkat cell lysate, HEK-293 cell lysate, Human brain tissue lysate, Mouse brain tissue lysate, Mouse heart tissue lysate, Rat brain tissue lysate, Rat heart tissue lysate, rat brain tissue, PC-12. |
| Subcellular location: | Cytoplasm, cytosol, Golgi apparatus, Endomembrane system, Mitochondrion outer membrane, Peroxisome, Membrane, clathrin-coated pit, Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane. |
| Recommended Dilutions:
WB IHC-P IF-Tissue IF-Cell FC |
1:1,000-1:5,000 1:10,000 1:500-1:2,000 1:100 1:1,000 |
| Uniprot #: | SwissProt: O00429 Human | Q8K1M6 Mouse | O35303 Rat |
| Alternative names: | DLP1 dnm1l DNM1L_HUMAN Dnm1p/Vps1p-like protein dnml1 DRP1 DVLP Dymple Dynamin 1 like Dynamin family member proline-rich carboxyl-terminal domain less Dynamin like protein Dynamin related protein 1 Dynamin-1-like protein Dynamin-like protein 4 Dynamin-like protein Dynamin-like protein IV Dynamin-related protein 1 DYNIV 11 EMPF EMPF1 FLJ41912 HdynIV VPS1 |
Images
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Fig1:
Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 100ng/mL nocodazole for 18 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 82 kDa Observed band size: 75 kDa Exposure time: Lane 1-4 (left): 1 minute 30 seconds; ECL: K1801; Exposure time: Lane 1-4 (right): 1 minute 40 seconds; ECL: K1802; 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 (HA722760) at 1/5,000 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 (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Flow cytometric analysis of HeLa cells labeling Phospho-DRP1 (S616). Cells were fixed and permeabilized. Then stained with the primary antibody (HA722760, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig3:
Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: C2C12 cell lysate Lane 3: PC-12 cell lysate Lane 4: HeLa cell lysate, the membrane blocked with phospho-peptide Lane 5: C2C12 cell lysate, the membrane blocked with phospho-peptide Lane 6: PC-12 cell lysate, the membrane blocked with phospho-peptide Lane 7: HeLa cell lysate, the membrane blocked with non-phospho-peptide Lane 8: C2C12 cell lysate, the membrane blocked with non-phospho-peptide Lane 9: PC-12 cell lysate, the membrane blocked with non-phospho-peptide Lysates/proteins at 20 µg/Lane. Predicted band size: 82 kDa Observed band size: 75 kDa Exposure time: 25 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 (HA722760) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/10,000 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 (HA722760) at 1/10,000 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. |
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Fig5:
Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: Jurkat cell lysate Lane 3: HEK-293 cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: C2C12 cell lysate Lane 6: PC-12 cell lysate Lane 7: Human brain tissue lysate Lane 8: Mouse brain tissue lysate Lane 9: Mouse heart tissue lysate Lane 10: Rat brain tissue lysate Lane 11: Rat heart tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 82 kDa Observed band size: 75 kDa Exposure time: 20 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 (HA722760) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/10,000 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 (HA722760) at 1/10,000 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. |
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Fig7:
Immunocytochemistry analysis of HeLa cells labeling Phospho-DRP1 (S616) with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) 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-Phospho-DRP1 (S616) antibody (HA722760) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig8:
Immunocytochemistry analysis of PC-12 cells labeling Phospho-DRP1 (S616) with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) 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-Phospho-DRP1 (S616) antibody (HA722760) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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