mAbs Against HUMAN Protein
RAF1
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | Rat monoclonal |
| CLONE NAME: | EMI411E |
| PROTEIN: | RAF proto-oncogene serine/threonine-protein kinase |
| PROTEIN WEB: | https://www.uniprot.org/uniprot/P04049 |
| ANTIGEN USED: | His-RAF1 (fragment 189-353) |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG2a |
| SPECIES REACTIVITY: | human and mouse |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
Description
Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2).
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | EMI411E | Neat | supernatant | / | |||||
| Paraffin tissue | |||||||||
| Immunofluorescence | |||||||||
Enlarge image
- EMI411E mAb is able to detect human and mouse RAF1 protein in immunocytochemistry
- To confirm that EMI411E mAb recognizes human and mouse RAF1 protein and do not cross react with mA-RAF and m/hBRAF, immunocytochemistry on frozen cytospin preparations of human and mouse proteins expressed in HEK293T were performed. Anti-FLAG, RAF1, B-RAF and V5 Abs were used as positive controls.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | EMI411E | Neat | supernatant | / | 73kDa | 73kDa | Available | |
| Immunoprecipitation | ||||||||
Enlarge image
- EMI411E mAb is able to detect mouse and human RAF1 protein by WB.
- LANES
Lane 1 HEK-mARAF (10ug) (-)
Lane 2 HEK-mBRAF (10ug) (-)
Lane 3 HEK-hBRAF (10ug) (-)
Lane 4 HEK-mRAF1 (10ug) (+)
Lane 5 HEK-hRAF1 (10ug) (+)
GAPDH was used as loading control
Enlarge image
- EMI411E mAb is able to detect mouse and human RAF1 protein by WB.
- LANES
Lane 1 mouse Lung Adenocarcinoma RAF1 KO (20ug) (-)
Lane 2 mouse Lung Adenocarcinoma RAF1 WT (20ug) (+)
Lane 3 human A549 (RAF1 WT) (20ug) (+)
Lane 4 human PULM24 (RAF1 WT) (20ug) (+)
Predicted molecular weight: 73kDa
Observed molecular weight: 73kDa