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.
Key Product Details
Species Reactivity
Human
Applications
Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2B Clone # 684128
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Product Specifications
Immunogen
E. coli-derived recombinant human A-Raf
Asn150-Lys314
Accession # P10398
Asn150-Lys314
Accession # P10398
Specificity
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.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2B
Scientific Data Images for Human A‑Raf Antibody
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.Applications for Human A‑Raf Antibody
Application
Recommended Usage
Western Blot
0.5 µg/mL
Sample: 293T human embryonic kidney cell line and HeLa human cervical epithelial carcinoma cell line
Sample: 293T human embryonic kidney cell line and HeLa human cervical epithelial carcinoma cell line
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: A-Raf
Long Name
v-Raf Murine Sarcoma 3611 Viral Oncogene Homolog
Alternate Names
ARaf, PKS, PKS2, RAFA1
Entrez Gene IDs
369 (Human)
Gene Symbol
ARAF
UniProt
Additional A-Raf Products
Product Documents for Human A‑Raf Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human A‑Raf Antibody
For research use only
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
IL-9 Signaling Pathways
IL-15 Signaling Pathways
IL-21 Signaling Pathways
MAPK Signaling: Oxidative Stress Pathway
MAPK Signaling Pathway: Mitogen Stimulation Pathway
mTOR Signaling Pathway
TGF-beta Signaling Pathways
VEGF - VEGF R2 Signaling Pathways