Antibody data
- Product number
- PA1-4639
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Phospho-c-Raf (Ser301) Polyclonal Antibody
- Provider product page
- Invitrogen Antibodies - PA1-4639
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with bovine, canine, chicken, mouse, non-human primate and Xenopus based on 100% sequence homology. This antibody is specific for the ~74 kDa Raf-1 protein phosphorylated at Ser301. Immunolabeling can be blocked by lambda-phosphatase treatment.
- Reactivity
- Human, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- Lot Dependent
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Supportive validation
- Submitted by
-
Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot of UV treated human Jurkat cell lysate showing specific immunolabeling of the ~74 kDa Raf-1 protein (Lane 1). The phosphospecificty of this labeling is shown in Lane 2 (lambda-phosphatase). The blot is identical to the control except that it was incubated in lambda-Ptase (1200 units for 30 min) before being exposed to the Anti-Ser301 Raf-1. The immunolabeling of Raf-1 is completely eliminated by treatment with lambda-Ptase.
Supportive validation
- Submitted by
-
Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with TPA (200 nM for 30 minutes) (Lane 2), COS-7 (Lane 3), COS-7 treated with TPA (200 nM for 30 minutes) (Lane 4), T-47D (Lane 5), T-47D treated with TPA (200 nM for 30 minutes) (Lane 6), NIH/3T3 (Lane 7) and NIH/3T3 treated with TPA (200 nM for 30 minutes) (Lane 8). The blot was probed with Anti-Phospho-RAF1 (Ser301) Rabbit Polyclonal Antibody (Product # PA1-4639, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 74 kDa band corresponding to Phospho-RAF1 (Ser301) was detected and observed to increase upon TPA treatment across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane using iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
