AHP1237 recognises Raf-1 phosphorylated at serine 642, a ubiquitously expressed 74 kDa member of the serine/threonine protein kinase family. Raf-1 is a MAP kinase kinase kinase and participates in the transduction of mitogenic signals from receptors on the cell membrane to the nucleus. Raf-1 is activated through phosphorylation by Ras and in turn phosphorylates MEK which activates ERK. ERK when activated regulates the expression of genes involved in cell cycle, differentiation, migration and apoptosis. As such Raf-1 is an important clinical target in cancer.
Raf-1 activity can be downregulated by phosphorylation at serine residues 301 and 642.
Species Cross-Reactivity
Target Species
Cross Reactivity
Bovine
Expected from Sequence
Chicken
Expected from Sequence
Dog
Expected from Sequence
Human
Expected from Sequence
Mouse
Expected from Sequence
Primate
Expected from Sequence
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
Flow Cytometry
Immunohistology - Frozen
Immunohistology - Paraffin
Immunoprecipitation
Western Blotting(1)
1/1000
(1)
For the detection of phosphoproteins, tyrosine phosphatase inhibitors such as 1mM Sodium Orthovanadate should be added to the sample buffer. Milk or other casein-based blocking solutions are not recommended as casein is a phosphoprotein and its use can result in high background.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Western Blotting
AHP1237 detects a band of approximately 74kDa.
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
12 months from date of despatch.
Antiserum Preparation
Antisera to rat Raf-1 (pSer 642) were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG prepared affinity chromatography.
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