Detection of Human, Mouse, and Rat Raf‑1 by Western Blot.
Western blot shows lysates of MCF‑7 human breast cancer cell line, HepG2 human hepatocellular carcinoma cell line, A431 human epithelial carcinoma cell line, L‑929 mouse fibroblast cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat Raf‑1 Monoclonal Antibody (Catalog # MAB4540) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Raf‑1 at approximately 74 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Raf‑1 in Human Liver.
Raf‑1 was detected in immersion fixed paraffin-embedded sections of human liver array using Mouse Anti-Human/Mouse/Rat Raf‑1 Monoclonal Antibody (Catalog # MAB4540) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
The Raf serine/threonine kinases are effectors of Ras that function as MAP3Ks in the ERK phosphorylation cascade. Mammals express three Raf proteins: A-Raf, B-Raf, and Raf-1, also known as C-Raf. Human Raf-1 contains three distinct regions; an N-terminal RBD (Ras-binding domain) (aa 56 - 131), followed by two rich segments [a cysteine-finger region (aa 138 - 184) (also called CR1/C1) and a second cysteine-rich region (CR2) (aa 253 - 264)] and a C-terminal Ser/Thr kinase catalytic domain (aa 354 - 611). Active Raf-1 phosphorylates and activates the MAPK kinases MEK1 and 2, which in turn phosphorylate and activate the MAP kinases ERK1 and 2.
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