WB result of c-Raf Recombinant Rabbit mAb
Primary antibody: c-Raf Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: A549 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 73 kDa
Observed MW: 68 kDa
c-Raf Recombinant Rabbit mAb (S-2718-61)
c-Raf Recombinant Rabbit mAb (S-2718-61)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | c-Raf |
| Synonyms | RAF proto-oncogene serine/threonine-protein kinase; Proto-oncogene c-RAF (cRaf); Raf-1; RAF; RAF1 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus, Mitochondrion, Cell membrane |
| Accession | P04049 |
| Clone Number | S-2718-61 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC |
| Reactivity | Hu, Ms, Rt, Mk |
| Positive Sample | HeLa, A549, C2C12, NIH/3T3, PC-12, COS-7 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000-1:2000 | Hu, Ms, Rt, Mk |
| ICC | 1:100 | Hu |
Background
c-Raf (also called Raf-1) is a 648-amino-acid serine/threonine kinase that serves as the pivotal upstream cytoplasmic MAP3K in the classical Ras→Raf→MEK→ERK/MAPK cascade: upon growth-factor- or oncogenic Ras-GTP-mediated recruitment to the plasma membrane, it homo- or hetero-dimerizes, becomes phosphorylated on multiple activating residues (Ser338, Tyr341, Thr491, Ser494) while relieving N-terminal autoinhibition, and then processively phosphorylates MEK1/2 to trigger downstream proliferative, anti-apoptotic, and differentiative transcriptional programs; its activity is tightly controlled via feedback and scaffold proteins, and gain-of-function mutations, overexpression, or dysregulated upstream signaling of c-Raf are implicated in numerous cancers, making it a key therapeutic target.
Picture
Picture
Western Blot
WB result of c-Raf Recombinant Rabbit mAb
Primary antibody: c-Raf Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: C2C12 whole cell lysate 20 µg
Lane 2: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 73 kDa
Observed MW: 68 kDa
WB result of c-Raf Recombinant Rabbit mAb
Primary antibody: c-Raf Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 73 kDa
Observed MW: 68 kDa
WB result of c-Raf Recombinant Rabbit mAb
Primary antibody: c-Raf Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 73 kDa
Observed MW: 68 kDa
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti-c-Raf antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
