Western blot analysis of SRC1 was performed by loading 25 ug of LNCaP (lane 1), PC-3 (lane 2) and NIH-3T3 (lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a SRC1 monoclonal antibody (Product # MA1-840) at a dilution of 1:2000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~152 and ~157 kDa.
|Tested species reactivity||Human, Mouse, Non-human primate|
|Published species reactivity||Rat, Non-human primate, Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to residues 477-947 of human SRC-1.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||5µl per 10^6 cells|
|Flow Cytometry (Flow)||1µg per 10^6 cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-840 detects steroid receptor coactivator-1 (SRC-1) from mouse and primate tissues and cells as well as recombinant human SRC-1 protein.
MA1-840 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot MA1-840 detects an ~165 kDa protein representing SRC-1 in COS cell extract. Also by Western blot, MA1-840 detects an unidentified protein at ~30-40 kDa. This antibody does not react with NIH-3T3 cells in Western blot.
The MA1-840 immunogen is a synthetic peptide corresponding to residues 477-947 of human SRC-1. The epitope for MA1-840 has been mapped to the extreme C-terminal end of this fragment.
Steroid and thyroid hormones and retinoic acid regulate a complex array of gene expression activity via intracellular receptor transcription factors belonging to the ligand dependent nuclear receptor superfamily. Adding to the complexity of function of these transcription factors are associated proteins known as coactivators and corepressors which, as their names suggest, enhance or depress transcriptional activity of the nuclear receptor with which they associate. One such coactivator is Steroid Receptor Coactivator-1 (SRC-1).
SRC-1 has been shown to interact with and stimulate the transcriptional activity of estrogen, progesterone, retinoic acid, thyroid and glucocorticoid receptors, as well as retinoic X receptor (RXR) in a ligand dependent manner. The amino terminal region of SRC-1 contains a PAS-A-basic helix-loop-helix homology domain which has previously been shown to be dimerization domains in other nuclear transcription factors including the aryl hydrocarbon (Ah) receptor and its heterodimerization partner, ARNT. The carboxy-terminal region of SRC-1 has been shown to possess histone acetyltransferase activity specific primarily for histones H3 and H4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
SIP1/NHERF2 enhances estrogen receptor alpha transactivation in breast cancer cells.
MA1-840 was used in immunoprecipitation and western blot to study the role of NHERF2 as an ER-alpha co-activator in breast cancer cells
|Meneses-Morales I,Tecalco-Cruz AC,Barrios-García T,Gómez-Romero V,Trujillo-González I,Reyes-Carmona S,García-Zepeda E,Méndez-Enríquez E,Cervantes-Roldán R,Pérez-Sánchez V,Recillas-Targa F,Mohar-Betancourt A,León-Del-Río A||Nucleic acids research (42:6885)||2014|
D538G mutation in estrogen receptor-¿: A novel mechanism for acquired endocrine resistance in breast cancer.
MA1-840 was used in immunoprecipitation to study the role of ER-alpha D538G mutation in mediating aquired resistance to hormone therapy in breast cancer patients
|Merenbakh-Lamin K,Ben-Baruch N,Yeheskel A,Dvir A,Soussan-Gutman L,Jeselsohn R,Yelensky R,Brown M,Miller VA,Sarid D,Rizel S,Klein B,Rubinek T,Wolf I||Cancer research (73:6856)||2013|
Expression patterns of the steroid receptor coactivator family in human ovarian endometriosis.
MA1-840 was used in immunohistochemistry to characterize the expression of estrogen and steroid receptor cofactors in human endometriosis
|Kumagami A,Ito A,Yoshida-Komiya H,Fujimori K,Sato A||The journal of obstetrics and gynaecology research (37:1269)||2011|
Role of SRC-1 in the promotion of prostate cancer cell growth and tumor progression.
MA1-840 was used in immunohistochemistry to study the role of SRC-1 in prostate cancer cell growth and tumor progression.
|Agoulnik IU,Vaid A,Bingman WE,Erdeme H,Frolov A,Smith CL,Ayala G,Ittmann MM,Weigel NL||Cancer research (65:7959)||2005|
Constitutive activation of the mitogen-activated protein kinase pathway impairs vitamin D signaling in human prostate epithelial cells.
MA1-840 was used in western blot to investigate the effect of constitutive activation of MAPK signaling on vitamin D signaling in immortalized human prostate epithelial cells
|Zhang Z,Kovalenko P,Cui M,Desmet M,Clinton SK,Fleet JC||Journal of cellular physiology (224:433)||2010|
Role of steroid receptor coactivators in glucocorticoid and transforming growth factor beta regulation of plasminogen activator inhibitor gene expression.
MA1-840 was used in western blot to study the regulation of plasminogen activator inhibitor gene expression through steroid receptor coactivators.
|Li G,Heaton JH,Gelehrter TD||Molecular endocrinology (Baltimore, Md.) (20:1025)||2006|
Ordered recruitment of histone acetyltransferases and the TRAP/Mediator complex to thyroid hormone-responsive promoters in vivo.
MA1-840 was used in western blot to study the role of histone acetylation in TRAP/Mediator complex recruitment and function.
|Sharma D,Fondell JD||Proceedings of the National Academy of Sciences of the United States of America (99:7934)||2002|
Endogenously expressed estrogen receptor and coactivator AIB1 interact in MCF-7 human breast cancer cells.
MA1-840 was used in western blot to demonstrate the role of AIB1 in human estrogen receptor signal pathway in MCF-7 human breast cancer cells.
|Tikkanen MK,Carter DJ,Harris AM,Le HM,Azorsa DO,Meltzer PS,Murdoch FE||Proceedings of the National Academy of Sciences of the United States of America (97:12536)||2000|
|Non-human primate||Not Cited||
Recruitment of nuclear receptor corepressor and coactivator to the retinoic acid receptor by retinoid ligands. Influence of DNA-heterodimer interactions.
MA1-840 was used in western blot to study the activation mechanism for retinoic acid receptors by their ligands.
|Klein ES,Wang JW,Khalifa B,Gavigan SA,Chandraratna RA||The Journal of biological chemistry (275:19401)||2000|
Partial hormone resistance in mice with disruption of the steroid receptor coactivator-1 (SRC-1) gene.
MA1-840 was used in western blot to investigate the functional properties of SRC-1 in mice
|Xu J,Qiu Y,DeMayo FJ,Tsai SY,Tsai MJ,O'Malley BW||Science (New York, N.Y.) (279:1922)||1998|
Steroid receptor coactivator-1 is a histone acetyltransferase.
MA1-840 was used in western blot to characterize the steroid receptor coactivator 1
|Spencer TE,Jenster G,Burcin MM,Allis CD,Zhou J,Mizzen CA,McKenna NJ,Onate SA,Tsai SY,Tsai MJ,O'Malley BW||Nature (389:194)||1997|
Nuclear coactivator-62 kDa/Ski-interacting protein is a nuclear matrix-associated coactivator that may couple vitamin D receptor-mediated transcription and RNA splicing.
MA1-840 was used in chromatin immunoprecipitation to investigate the role of nuclear coactivator-62 kDa/Ski-interacting protein (NCoA62/SKIP) during the vitamin D receptor (VDR)-mediated transcription and RNA splicing.
|Zhang C,Dowd DR,Staal A,Gu C,Lian JB,van Wijnen AJ,Stein GS,MacDonald PN||The Journal of biological chemistry (278:35325)||2003|