AHP1342 detects the p65 (RelA) subunit of nuclear factor kappa-B (NFkB) when phosphorylated at serine 536. NFkB is a heterodimeric DNA binding protein which was originally identified consisting of p65 and p50 (NFkB1) subunits. Research has led to the discovery of other subunits, including p52 (NFkB2), cRel, and RelB.
NFkB is ubiquitously expressed, and functions as a second messenger which upon activation leads to the transcription of a number of genes. NFkB is activated by a number of agents, including pro-inflammatory cytokines and bacterial lipopolysaccharide, and is essential for the maturation of T cells and regulation of their survival and activation.
Phosphorylation of serine 536 contributes to NFkB transactivation and increases p65 transcription activity.
Species Cross-Reactivity
Target Species
Cross Reactivity
Mouse
Expected from Sequence
Rat
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
1/500 - 1/3000
Flow Cytometry
Immunohistology - Frozen
Immunohistology - Paraffin
1/50 - 1/250
Immunoprecipitation
Western Blotting(1)
1/100 - 1/1000
(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 antibody 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 antibody for use in their own system using appropriate negative/positive controls.
Western Blotting
AHP1342 detects a band of approximately 65kDa in the appropriate cell lysate.
Storage
Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost free freezers is not recommended. 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 NFkB p65 (pSer536) were raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
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