Human/Mouse/Rat Raf-1 Antibody Summary
Asn189-Thr353
Accession # P04049
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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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.
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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.
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Detection of Human Raf‑1 by Simple WesternTM. Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Raf‑1 at approximately 75 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat Raf‑1 Monoclonal Antibody (Catalog # MAB4540). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Reconstitution Calculator
Preparation and Storage
- 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.
Background: Raf-1
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.
Product Datasheets
Citations for Human/Mouse/Rat Raf-1 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 2
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Oncogenic RAS drives the CRAF‐dependent extracellular vesicle uptake mechanism coupled with metastasis
Authors: Dongsic Choi, Laura Montermini, Brian Meehan, Anthoula Lazaris, Peter Metrakos, Janusz Rak
Journal of Extracellular Vesicles
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Macrocyclic lactones inhibit nasopharyngeal carcinoma cells proliferation through PAK1 inhibition and reduce in vivo tumor growth
Authors: F Gallardo, B Mariamé, R Gence, AF Tilkin-Mar
Drug Des Devel Ther, 2018-09-07;12(0):2805-2814.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
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30 ug of protein per sample.
1:500 dilution of primary Ab.
