Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3433(2)] to Raf1 (phospho S259)
- Suitable for: WB, Dot blot
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Raf1 (phospho S259) antibody [EPR3433(2)]
See all Raf1 primary antibodies -
Description
Rabbit monoclonal [EPR3433(2)] to Raf1 (phospho S259) -
Host species
Rabbit -
Specificity
This antibody only detects Raf1 when phosphorylated at Serine 259.
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Tested Applications & Species
See all applications and species dataApplication Species WB MouseRatHuman -
Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Raf1 aa 250-350 (phospho S259) (Cysteine residue). The exact sequence is proprietary.
Database link: P04049 -
Positive control
- WB: Treated C6, NIH/3T3 and HEK-293 whole cell lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3433(2) -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/5000 dilution (Purified)
Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour whole cell lysates
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour whole cell lysates, then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa
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All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution (Purified)
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 5uM forskolin for 10 minutes whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 5uM forskolin for 10 minutes whole cell lysates, then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa
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Lane 1 : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution (Purified)
Lanes 2-3 : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysates
Lane 2 : C6 (Rat glial tumor glial cell) treated with 30uM forskolin for 20 minutes whole cell lysates
Lane 3 : C6 (Rat glial tumor glial cell) treated with 30uM forskolin for 20 minutes whole cell lysates, then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa
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All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/5000 dilution
Lane 1 : HEK293 whole cell lysate
Lane 2 : HEK293 whole cell lysate treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour
Lane 3 : HEK293 whole cell lysate treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa
Exposure time: 1 secondBlocking and dilution buffer: 5% NFDM/TBST.
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Dot blot analysis of Raf1 (pS259) peptide (Lane 1) and Raf1 non-phospho peptide (Lane 2) labelling Raf1 (pS259) with ab173539 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution
Lane 1 : 293T cell lysates
Lane 2 : 293T cell lysates treated with Okadaic Acid + Calyculin A
Lysates/proteins at 10 µg per lane.
Predicted band size: 73 kDa
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