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Phosphosolutions.com

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Anti-Phospho-Ser294 Progesterone Receptor

Catalog Number: p205-294 Size: 100 µl Clone: 608 Subclass: IgG1
Product Description: Affinity purified mouse monoclonal antibody
Applications:
WB: 1:1000
IHC (frozen sections; unpublished observations): 1:1000
Antigen: Phosphopeptide corresponding to amino acid residues surrounding the phospho-
Ser294 of human progesterone receptor.

Species reactivity
: The antibody has been directly tested for reactivity in Western blots with
human tissue. It is anticipated that the antibody will also react with non-human primates based
on the fact that these species have 100% homology with the amino acid sequence used as
antigen.

Biological Significance:
There is accumulating evidence to suggest that progesterone plays
an essential role in the regulation of growth and differentiation of mammary glands and thus may
play a key role in breast cancer (Edwards, 2005). The biological response to progesterone is
mediated by two distinct forms of the human progesterone receptor (PR-A and PR-B forms). In
most cell contexts, the B form functions as a transcriptional activator, whereas the A form
functions as a transcriptional inhibitor of steroid hormones (Attia et al., 2000; Lin et al., 2003).
Recently it has been demonstrated that there is differential hormone dependent regulation of the
phosphorylation of the A and B forms of the receptor (Clemm et al., 2000) . Treatment of T47D
breast cancer cells with progestin agonist increases the phosphorylation of Ser190 and Ser294
with different kinetics. These phosphorylation events may differentially affect the transcriptional
activity of the receptor.
Anti-Phospho Ser294 Progesterone Receptor
Western blot of whole cell T47D lysate prepared from cells that had been incubated in
the presence of the synthetic progestin agonist R5020 (500 nM) showing specific
immunolabeling of the ~90k PR-A isoform and the ~120 PR-B isoform of the
progesterone receptor phosphorylated at Ser294. The immunolabeling is blocked by the
phosphopeptide used as the antigen (not shown).

WB
= Western Blot IF = Immunofluorescence IHC = Immunohistochemistry IP = Immunoprecipitation
Packaging:
100 µl in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 µg per ml BSA and 50% glycerol. Adequate amount of material to conduct
10-mini Western Blots.
Storage and Stability. For long term storage –20oC is recommended. Stable at –20oC for at least 1 year.
Shipment: Domestic - Blue Ice; International – Blue Ice or Dry Ice.
Purification Method: Prepared by affinity purification using a protein G column.

Antibody Specificity:
Specific for the ~90k PR-A isoform and the ~120k PR-B isoform
phosphorylated at Ser294. Immunolabeling is blocked by the phosphopeptide used as the
antigen but not by the corresponding dephosphopeptide.

Quality Control Tests:
Western blots performed on each lot.
References:
Attia GR, Zeitoun K, Edwards D, Johns A, Carr BR, Bulun SE (2000) Progesterone receptor isoform A but not B is expressed in
endometriosis. J Clin Endocrinol Metab 85:2897-2902. Clemm DL, Sherman L, Boonyaratanakornkit V, Schrader WT, Weigel NL, Edwards DP (2000) Differential hormone-dependent phosphorylation of progesterone receptor A and B forms revealed by a phosphoserine site-specific monoclonal antibody. Mol Endocrinol 14:52-65. Edwards DP (2005) Regulation of signal transduction pathways by estrogen and progesterone. Annu Rev Physiol 67:335-376. Lin VC, Woon CT, Aw SE, Guo C (2003) Distinct molecular pathways mediate progesterone-induced growth inhibition and focal adhesion. Endocrinology 144:5650-5657.
WB
= Western Blot IF = Immunofluorescence IHC = Immunohistochemistry IP = Immunoprecipitation
Packaging:
100 µl in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 µg per ml BSA and 50% glycerol. Adequate amount of material to conduct
10-mini Western Blots.
Storage and Stability. For long term storage –20oC is recommended. Stable at –20oC for at least 1 year.
Shipment: Domestic - Blue Ice; International – Blue Ice or Dry Ice.

Source: http://www.phosphosolutions.com/store/docs/26_datasheet.pdf

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