Detects human A-Raf in direct ELISAs and Western blots.
In direct ELISAs, no cross-reactivity with recombinant human (rh) B-Raf, rhRaf-1 (aa 1-648), rhRaf-1 (aa 189-353), or rhKSR1 is observed. In Western blots, no cross-reactivity
with recombinant human (rh) B-Raf or rhKSR1 is observed.
Monoclonal Mouse IgG2B Clone # 684128
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human A-Raf Asn150-Lys314 Accession # P10398
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
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.
Detection of Human A‑Raf by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line and HeLa human cervical epithelial carcinoma cell line. PVDF Membrane was probed with 0.5 µg/mL of Human A‑Raf Monoclonal Antibody (Catalog # MAB6607) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for A‑Raf at approximately 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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 A-Raf is a 70 kDa protein that shares three regions with B-Raf and Raf-1: aa 14-153 which contains a Ras-binding domain, a Cys-rich domain, and a lipid-binding domain; aa 209-223; and aa 308-573 which contains the Ser/Thr protein kinase domain and a second lipid-binding domain. A-Raf is activated by phosphorylation at Ser 257, 262, and 264. It then regulates multiple processes including endocytic trafficking, glycolysis, cell cycle progression, and apoptosis. Within aa 150-314, human A-Raf shares 87% aa sequence identity with mouse and rat A-Raf.
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