Product Pathways - Nuclear Receptor Signaling
Progesterone Receptor Signaling Antibody Sampler Kit #12807
|12807S||1 Kit (6 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370||40 µl||W, IP, IHC-P, IF-IC, F||H, M, R, Hm, Mk, Mi, Dm, Z, B, Dg, Pg, Sc||C, Ce||44, 42||Rabbit IgG|
|Phospho-Progesterone Receptor (Ser190) Antibody #3171||40 µl||W, IP||H||90, 118||Rabbit|
|Phospho-Progesterone Receptor (Ser345) Antibody #12783||40 µl||W||H||90 (PR-A), 118 (PR-B)||Rabbit|
|Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb #8757||40 µl||W, IP, IHC-P, IF-IC, ChIP||H||Mk||90 (PR-A), 118 (PR-B)||Rabbit IgG|
|Progesterone Receptor B (C1A2) Rabbit mAb #3157||40 µl||W, IHC-P, IF-IC, F||H||118||Rabbit IgG|
|Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb #6943||40 µl||W, IP||H, M, R||60||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
Reactivity Key: H=Human, M=Mouse, R=Rat, Hm=Hamster, Mk=Monkey, Mi=Mink, Dm=D. melanogaster, Z=Zebrafish, B=Bovine, Dg=Dog, Pg=Pig, Sc=S. cerevisiae
Western blot analysis of extracts from T-47D cells grown for 48 hr in phenol red-free medium supplemented with 5% charcoal-stripped FBS, untreated (-) or promegestone (R5020)-treated (100 nM, 1 hr; +), using Phospho-Progesterone Receptor (Ser345) Antibody #12783 (upper) or Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb #8757 (lower).
Western blot analysis of extracts from MCF7 and T-47D cells using Progesterone Receptor B (C1A2) Rabbit mAb #3157.
Western blot analysis of extracts from T-47D cells, untreated (-) or promegestone (R5020)-treated (100 nm, 1 hr; +), using Phospho-Progesterone Receptor (Ser190) Antibody #3171 (upper) and control Progesterone Receptor Antibody #3172 (lower).
Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA #4174 as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).
Western blot analysis of extracts NIH/3T3 cells, serum-starved (-) or treated with human Platelet-Derived Growth Factor BB hPDGF-BB #8912 (100 ng/ml, 15 min; +), using Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb #6943 (upper) or Src (36D10) Rabbit mAb #2109 (lower).
Western blot analysis of extracts from T-47D (PR positive) and MDA-MB-231 (PR negative) cells using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb #8757 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Progesterone Receptor Signaling Antibody Sampler Kit provides an economical means of detecting total and active levels of progesterone receptor (PR) as well as the active forms of PR downstream targets. The kit contains enough primary antibody to perform four western blots per primary antibody.
Specificity / Sensitivity
Unless otherwise indicated, each antibody will recognize endogenous total levels of target protein. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb recognizes endogenous p44 and p42 MAP Kinase (Erk1/2) when dually phosphorylated at Thr202/Tyr204 of Erk1 (Thr185/Tyr187 of Erk2), and singly phosphorylated at Thr202. This antibody does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinases. Phospho-Progesterone Receptor (Ser190) Antibody detects endogenous levels of progesterone receptors B and A only when phosphorylated at Ser190 and Ser26, respectively. Phospho-Progesterone Receptor (Ser345) Antibody recognizes endogenous levels of progesterone receptors B and A only when phosphorylated at Ser345 and Ser181, respectively. Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb may cross-react with other Src family proteins phosphorylated at equivalent sites or with overexpressed phosphorylated RTKs.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase, or with synthetic peptides corresponding to residues surrounding Ser115 or Tyr541 of human progesterone receptor protein, or Tyr416 of human Src protein. Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser190 of human progesterone receptor protein or Ser345 of human progesterone receptor B protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Human progesterone receptor (PR) is expressed as two forms: the full length PR-B and the short form PR-A. PR-A lacks the first 164 amino acid residues of PR-B (1,2). Both PR-A and PR-B are ligand activated, but differ in their relative ability to activate target gene transcription (3,4). The activity of PR is regulated by phosphorylation; at least seven serine residues are phosphorylated in its amino-terminal domain. Three sites (Ser81, Ser102, and Ser162) are unique to full length PR-B, while other sites (Ser190, Ser294, Ser345, and Ser400) are shared by both isoforms (5). Phosphorylation of PR-B at Ser190 (equivalent to Ser26 of PR-A) is catalyzed by CDK2 (6). Mutation of Ser190 results in decreased activity of PR (7), suggesting that the phosphorylation at Ser190 may be critical to its biological function. Research studies have demonstrated ligand-dependent phosphorylation of PR-B at Ser345 is catalyzed by MAPK and plays an important role in mediating the proliferation of breast cancer cells. Investigators have shown that Ser345-phosphorylated PR-B associates with Sp1 to regulate EGFR and p21 transcription (8). PR signaling has been shown to crosstalk with other kinase signaling cascades. Upon stimulation and the subsequent interaction with estrogen receptor α and c-Src, PR-B has been shown to promote the activation of the Src/p21ras/Erk pathway (9).
- Evans, R.M. (1988) Science 240, 889-95.
- Kastner, P. et al. (1990) EMBO J 9, 1603-14.
- Giangrande, P.H. et al. (2000) Mol Cell Biol 20, 3102-15.
- Wen, D.X. et al. (1994) Mol Cell Biol 14, 8356-64.
- Clemm, D.L. et al. (2000) Mol Endocrinol 14, 52-65.
- Zhang, Y. et al. (1997) Mol Endocrinol 11, 823-32.
- Takimoto, G.S. et al. (1996) J Biol Chem 271, 13308-16.
- Faivre, E.J. et al. (2008) Mol Endocrinol 22, 823-37.
- Blunt, R.J. et al. (1975) Pflugers Arch 355, 189-204.
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* Product-specific protocol.
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 9998 BSA
- 9999 Nonfat Dry Milk
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 9997 Tris Buffered Saline with Tween® 20 (TBST-10X)
- 7722 Blue Loading Buffer Pack
- 7723 Red Loading Buffer Pack
For Research Use Only. Not For Use In Diagnostic Procedures.
XP® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
This antibody is developed, validated, and produced by CST using in part technology under license (granting certain rights including those under U.S. Patent No. 5,675,063) from Epitomics, Inc.