Product Pathways - Nuclear Receptor Signaling
Nuclear Receptor Antibody Sampler Kit #8595
|8595S||1 Kit (8 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|RARα Antibody #2554||40 µl||W||M, R||55||Rabbit|
|RARγ1 (D3A4) XP® Rabbit mAb #8965||40 µl||W, IP, IHC-P, IF-IC||H, M||R, Hm, B, Dg||58||Rabbit IgG|
|RXRα (D6H10) Rabbit mAb #3085||40 µl||W, IP||H, M, R||53||Rabbit IgG|
|Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660||40 µl||W, IP, IF-IC, F, ChIP||H, M, R, Mk||80, 91, 94||Rabbit IgG|
|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|
|Androgen Receptor (D6F11) XP® Rabbit mAb #5153||40 µl||W, IHC-P, IF-IC, F||H||110||Rabbit IgG|
|Estrogen Receptor α (D8H8) Rabbit mAb #8644||40 µl||W, IP, IF-IC, ChIP||H||66||Rabbit IgG|
|PPARγ (C26H12) Rabbit mAb #2435||40 µl||W, IHC-P, IF-IC||H, M||R||53, 57||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: M=Mouse, R=Rat, H=Human, Mk=Monkey
Western blot analysis of extracts from various cell lines using Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660.
Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells (differentiated 6 d) using PPARγ (C26H12) Rabbit mAb #2435.
Western blot analysis of extracts from AR-positive (LNCaP and MCF7) and AR-negative (PC-3 and DU 145) cell lines using Androgen Receptor (D6F11) XP® Rabbit mAb #5153 (upper) or β-Actin Antibody #4967 (lower).
Western blot analysis of extracts from ER-positive (MCF7, T-47D, ZR-75-1) and ER-negative (SK-BR-3 and MCF 10A) cell lines using Estrogen Receptor α (D8H8) Rabbit mAb #8644 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from PR-positive (T-47D) and PR-negative (MDA-MB-231) cell lines using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb #8757 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
The Nuclear Receptor Antibody Sampler Kit provides an economical means to evaluate the presence and status of nuclear receptors. This kit contains enough primary antibody to perform four western blots per primary.
Specificity / Sensitivity
Each antibody in the Nuclear Receptor Antibody Sampler Kit recognizes endogenous levels of total respective protein. Glucocorticoid Receptor (D8H2) XP® Rabbit mAb is predicted to cross-react with all known alternative translation start site generated isoforms of glucocorticoid receptor-α and glucocorticoid receptor-β, and does not cross-react with mineralocorticoid receptor. Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb does not cross-react with either the glucocorticoid receptor or the mineralocorticoid receptor. RARγ1 (D3A4) XP® Rabbit mAb is not predicted to cross-react with RARγ2, and does not cross-react with either RARα or RARβ. RXRα (D6H10) Rabbit mAb does not cross-react with either RXRβ or RXRγ.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human RARα protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminal region of human androgen receptor protein, residues in the carboxy terminus of human ERα protein, residues surrounding Leu378 of human glucocorticoid receptor protein, residues surrounding Asp69 of human PPARγ protein, residues surrounding Tyr541 of human progesterone receptor protein, residues near the amino terminus of human RARγ1 protein, or residues near the amino terminus of human RXRα protein.
Nuclear Receptors are transcription factors responsible for sensing bioactive molecules, including steroid and thyroid hormones. They are regulated by multiple posttranslational modifications, which in turn impacts their ability to regulate the expression of specific genes involved in the control of reproduction, development, and metabolism.
Androgen receptor (AR), a zinc finger transcription factor belonging to the nuclear receptor superfamily, is activated by phosphorylation and dimerization upon ligand binding (1). This promotes nuclear localization and binding of AR to androgen response elements in androgen target genes. AR plays a crucial role in several stages of male development and the progression of prostate cancer (2,3).
Estrogen receptor α (ERα), a member of the steroid receptor superfamily, contains highly conserved DNA binding and ligand binding domains (4). Through its estrogen-independent and estrogen-dependent activation domains (AF-1 and AF-2, respectively), ERα regulates transcription by recruiting coactivator proteins and interacting with general transcriptional machinery (5).
Glucocorticoid hormones control cellular proliferation, inflammation, and metabolism through their association with the glucocorticoid receptor (GR)/NR3C1, a member of the nuclear hormone receptor superfamily of transcription factors (6).
Peroxisome proliferator-activated receptor γ (PPARγ) is a member of the ligand-activated nuclear receptor superfamily and functions as a transcriptional activator (7). PPARγ is preferentially expressed in adipocytes, as well as in vascular smooth muscle cells and macrophages (8). Besides its role in mediating adipogenesis and lipid metabolism (8), PPARγ also modulates insulin sensitivity, cell proliferation, and inflammation (9).
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 (10,11). Both PR A and PR B are ligand activated, but differ in their relative ability to activate target gene transcription (12,13).
Nuclear retinoic acid receptors (RARs) consist of three subtypes encoded by separate genes: α (NR1B1), β (NR1B2), and γ (NR1B3). For each subtype, there are at least two isoforms, which are generated by differential promoter usage and alternative splicing and differ only in their N-terminal regions. Retinoids, which are metabolites of vitamin A, serve as ligands for RARs (14). RARs function as ligand-dependent transcriptional regulators and are found to be heterodimerized with retinoid X receptors (RXRs). These transcriptionally active dimers regulate the expression of genes involved in cellular differentiation, proliferation, and apoptosis (15,16).
The human retinoid X receptors are encoded by three distinct genes (RXRα, RXRβ, and RXRγ) and bind selectively and with high affinity to the vitamin A derivative, 9-cis-retinoic acid. RXRs are type-II nuclear hormone receptors that are largely localized to the nuclear compartment independent of ligand binding. Nuclear RXRs form heterodimers with nuclear hormone receptor subfamily 1 proteins, including thyroid hormone receptor, retinoic acid receptors, vitamin D receptor, peroxisome proliferator-activated receptors, liver X receptors, and farnesoid X receptor (17).
- Li, J. and Al-Azzawi, F. (2009) Maturitas 63, 142-8.
- Avila, D.M. et al. (2010) J Steroid Biochem Mol Biol 76, 135-42.
- Montgomery, J.S. et al. (2001) J Pathol 195, 138-46.
- Mangelsdorf, D.J. et al. (1995) Cell 83, 835-9.
- Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev 14, 121-41.
- Yamamoto, K.R. (1985) Annu Rev Genet 19, 209-52.
- Tontonoz, P. et al. (1995) Curr Opin Genet Dev 5, 571-6.
- Rosen, E.D. et al. (1999) Mol Cell 4, 611-7.
- Murphy, G.J. and Holder, J.C. (2000) Trends Pharmacol Sci 21, 469-74.
- 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.
- Rochette-Egly, C. and Germain, P. (2009) Nucl Recept Signal 7, e005.
- Delacroix, L. et al. (2010) Mol Cell Biol 30, 231-44.
- Eifert, C. et al. (2006) Mol Reprod Dev 73, 796-824.
- Gronemeyer, H. et al. (2004) Nat Rev Drug Discov 3, 950-64.
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.