Phospho-Raf1 (S259) Recombinant Rabbit Monoclonal Antibody [SD85-07]
cat.: ET1612-87
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | SD85-07 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 73 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser259 of Human Raf1 aa 236-285 / 648. |
| Positive control: | HEK-293 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, HEK-293, human skeletal muscle tissue, mouse skeletal muscle tissue, rat skeletal muscle tissue, human testis tissue, human tracheal tissue. |
| Subcellular location: | Cytoplasm, Cell membrane, Mitochondrion, Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:2,000 1:400-1:1,000 1:100 1:1,000 |
| Uniprot #: | SwissProt: P04049 Human | Q99N57 Mouse | P11345 Rat |
| Alternative names: | c Raf C-raf C-Raf proto-oncogene, serine/threonine kinase CMD1NN Craf 1 transforming gene cRaf Craf1 transforming gene EC 2.7.11.1 kinase Raf1 Murine sarcoma 3611 oncogene 1 NS5 Oncogene MIL Oncogene RAF1 OTTHUMP00000160218 OTTHUMP00000207813 OTTHUMP00000209389 Protein kinase raf 1 Proto-oncogene c-RAF Raf 1 Raf 1 proto oncogene serine/threonine kinase RAF Raf proto oncogene serine/threonine protein kinase RAF proto-oncogene serine/threonine-protein kinase RAF-1 RAF1 RAF1_HUMAN Similar to murine leukemia viral (V-raf-1) oncogene homolog 1 TRANSFORMING REPLICATION-DEFECTIVE MURINE RETROVIRUS 3611-MSV v raf 1 murine leukemia viral oncogene homolog 1 v-raf murine sarcoma viral oncogene homolog 1 v-raf-1 murine leukemia viral oncogene-like protein 1 vraf1 murine leukemia viral oncogene homolog 1 |
Images
|
Fig1:
Western blot analysis of Phospho-Raf1 (S259) on different lysates with Rabbit anti-Phospho-Raf1 (S259) antibody (ET1612-87) at 1/2,000 dilution. Lane 1: HEK-293 cell lysate Lane 2: HeLa cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: C6 cell lysate Lane 5: HEK-293 cell lysate, the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 73 kDa Observed band size: 65 kDa Exposure time: 59 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 (ET1612-87) at 1/2,000 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. |
|
Fig2:
Immunocytochemistry analysis of HEK-293 cells labeling Phospho-Raf1 (S259) with Rabbit anti-Phospho-Raf1 (S259) antibody (ET1612-87) 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-Phospho-Raf1 (S259) antibody (ET1612-87) 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 (HA601187, 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. |
|
Fig3:
Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-Phospho-Raf1 (S259) antibody (ET1612-87) 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 (ET1612-87) 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. |
|
Fig4:
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-Phospho-Raf1 (S259) antibody (ET1612-87) 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 (ET1612-87) 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. |
|
Fig5:
Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-Phospho-Raf1 (S259) antibody (ET1612-87) 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 (ET1612-87) 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. |
|
Fig6:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Phospho-Raf1 (S259) antibody (ET1612-87) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1612-87) at 1/500 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. |
|
Fig7:
Immunohistochemical analysis of paraffin-embedded human tracheal tissue with Rabbit anti-Phospho-Raf1 (S259) antibody (ET1612-87) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1612-87) at 1/400 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. |
|
Fig8:
Flow cytometric analysis of HEK-293 cells labeling Phospho-Raf1 (S259). Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-87, 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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.