AHP1333B detects phosphorylated threonine (phosphothreonine), an alpha amino acid that together with phosphoserine is one of the most ubiquitously modified amino acids in proteins. Many hormones, such as adrenaline (epinephrine), alter the activities of enzymes by stimulating the phosphorylation of serine and threonine, and these can have profound effects on cellular signal transduction, apoptosis and carcinogenesis.
AHP1333B detects phosphothreonine in multiple forms including free phosphothreonine, phosphothreonine conjugated to carriers such as thyroglobulin or BSA, and in proteins of both unstimulated and stimulated cell lysates.
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
1/5000
Flow Cytometry
Immunohistology - Frozen
Immunohistology - Paraffin
Immunoprecipitation
Western Blotting(1)
1/500
(1)
For the detection of phosphoproteins, threonine phosphatase inhibitors such as 10mM Sodium Fluoride 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.
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
18 months from date of despatch.
Antiserum Preparation
Antisera to phosphothreonine were raised by repeated immunisations of rabbits with highly purified antigen.
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