Revision 1

#84200Store at -20C

1 Kit

(5 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Androgen Receptor (D6F11) XP® Rabbit mAb 5153 20 µl 110 kDa Rabbit IgG
Estrogen Receptor α (D8H8) Rabbit mAb 8644 20 µl 66 kDa Rabbit IgG
Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb 12041 20 µl 94, 91 kDa Rabbit IgG
Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb 8757 20 µl 90 (PR-A), 118 (PR-B) kDa Rabbit IgG
Mineralocorticoid Receptor (E9W1M) Rabbit mAb 58883 20 µl 120 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Steroid Hormone Receptor Antibody Sampler Kit provides an economical means of detecting levels of steroid hormone nuclear receptors. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

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. Research studies have shown that 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). 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 (6,7). Both PR-A and PR-B are ligand activated, but differ in their relative ability to activate target gene transcription (8,9). 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 (10). GR is composed of several conserved structural elements, including a carboxy-terminal ligand-binding domain (which also contains residues critical for receptor dimerization and hormone-dependent gene transactivation), a neighboring hinge region containing nuclear localization signals, a central zinc-finger-containing DNA-binding domain, and an amino-terminal variable region that participates in ligand-independent gene transcription. In the absence of hormone, a significant population of GR is localized to the cytoplasm in an inactive form via its association with regulatory chaperone proteins, such as HSP90, HSP70, and FKBP52. On hormone binding, GR is released from the chaperone complex and translocates to the nucleus as a dimer to associate with specific DNA sequences termed glucocorticoid response elements (GREs), thereby enhancing or repressing transcription of specific target genes (11). Mineralocorticoid receptor (MR) is a steroid hormone receptor with structural and functional similarities to GR. MR binds with high affinity to aldosterone and other mineralocorticoids as well as glucocorticoids (12-14). Upon ligand binding, MR undergoes conformational changes and enters the nucleus to bind to target mineralocorticoid response elements (MREs) (4,15,16). MR is also able to heterodimerize with GR and bind to hormone response elements on DNA in cells that express both receptors (17-19). 

  1. Li, J. and Al-Azzawi, F. (2009) Maturitas 63, 142-8.
  2. Avila, D.M. et al. (2001) J Steroid Biochem Mol Biol 76, 135-42.
  3. Montgomery, J.S. et al. (2001) J Pathol 195, 138-46.
  4. Mangelsdorf, D.J. et al. (1995) Cell 83, 835-9.
  5. Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev 14, 121-41.
  6. Evans, R.M. (1988) Science 240, 889-95.
  7. Kastner, P. et al. (1990) EMBO J 9, 1603-14.
  8. Giangrande, P.H. et al. (2000) Mol Cell Biol 20, 3102-15.
  9. Wen, D.X. et al. (1994) Mol Cell Biol 14, 8356-64.
  10. Yamamoto, K.R. (1985) Annu Rev Genet 19, 209-52.
  11. Necela, B.M. and Cidlowski, J.A. (2003) Trends Pharmacol Sci 24, 58-61.
  12. Arriza, J.L. et al. (1987) Science 237, 268-75.
  13. Giguère, V. et al. (1988) Nature 331, 91-4.
  14. Beato, M. et al. (1995) Cell 83, 851-7.
  15. Guiochon-Mantel, A. et al. (1996) J Steroid Biochem Mol Biol 56, 3-9.
  16. Liu, W. et al. (1995) Proc Natl Acad Sci U S A 92, 12480-4.
  17. Liu, W. et al. (1996) Mol Endocrinol 10, 1399-406.
  18. Trapp, T. et al. (1994) Neuron 13, 1457-62.
  19. Funder, J.W. (1995) J Steroid Biochem Mol Biol 53, 53-5.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    For Research Use Only. Not for Use in Diagnostic Procedures.
    Revision 1
    #84200

    Steroid Hormone Receptor Antibody Sampler Kit

    Steroid Hormone Receptor Antibody Sampler Kit: Image 1 Expand Image
    CUT&RUN was performed with 2.5mg mouse liver tissue fixed with 0.1% formaldehyde for 10 minutes and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 8 (upper), including Chd9 (lower), a known target gene of Glucocorticoid Receptor (see additional figure containing CUT&RUN-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 2 Expand Image
    CUT&RUN was performed with 2.5mg mouse liver tissue fixed with 0.1% formaldehyde for 10 minutes and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Chd9, a known target gene of Glucocorticoid Receptor (see additional figure containing CUT&RUN-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 3 Expand Image
    CUT&RUN was performed with 2.5 mg mouse liver tissue fixed with 0.1% formaldehyde for 10 minutes and either Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using mouse A230028O05Rik promoter primers, mouse Chd9 Intron 2 primers and SimpleChIP® Mouse Intracisternal A-Particle (IAP) LTR Primers #85916. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from LNCaP (AR+), VCaP (AR & AR-V7+), 22Rv1 (AR & AR-V7+), and PC-3 (AR-) cells using Androgen Receptor (D6F11) XP® Rabbit mAb (upper), and β-Actin (13E5) Rabbit mAb (lower).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 5 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from LNCaP cells using Androgen Receptor (D6F11) XP® Rabbit mAb #5153. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 6 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from MCF7 cells using Estrogen Receptor α (D8H8) Rabbit mAb #8644. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 8 Expand Image
    Confocal immunofluorescent analysis of HeLa cells (left, positive) or HT-29 cells (right, negative) using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb #12041 (green), DyLight™ 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from LNCaP (AR+), MCF7 (AR+), PC-3 (AR-), and DU 145 (AR-) cells using Androgen Receptor (D6F11) XP® Rabbit mAb (upper) and β-Actin Antibody #4967 (lower).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 10 Expand Image
    CUT&RUN was performed with LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and Androgen Receptor (D6F11) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across KLK2, a known target gene of Androgen Receptor (see additional figure containing CUT&RUN-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from various cell lines using Mineralocorticoid Receptor (E9W1M) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, MSTO-211H is higher for mineralocorticoid receptor expression than in other cell lines.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 12 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 13 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and Estrogen Receptor α (D8H8) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across TFF1/pS2, a known target gene of Estrogen Receptor α (see additional figure containing ChIP-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from ER-positive cell lines (MCF7, T-47D, ZR-75-1) and ER-negative cell lines (SK-BR-3 and MCF 10A) using Estrogen Receptor α (D8H8) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 15 Expand Image
    CUT&RUN was performed with MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 days, then treated with β-estradiol (10 nM) for 45 minutes and either Estrogen Receptor a (D8H8) Rabbit mAb or Estrogen Receptor α (D6R2W) Rabbit mAb #13258, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® #56795. The figure shows binding across TFF1 gene.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 16 Expand Image
    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 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 17 Expand Image
    CUT&RUN was performed with T-47D cells cultured in phenol red-free media supplemented with 5% charcoal-stripped FBS for 48 hr and then treated with promegestone (R5020, 10 nM, 1 hr) and Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across SETD9 gene.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human glucocorticoid receptor-α (hGRα-Myc/DDK; +) or Myc/DDK-tagged full-length human mineralocorticoid receptor (hMR-Myc/DDK; +), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded LNCaP (AR+, left) and DU145 (AR-, right) using Androgen Receptor (D6F11) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 20 Expand Image
    CUT&RUN was performed with LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and Androgen Receptor (D6F11) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 19 (upper), including KLK2 (lower), a known target gene of Androgen Receptor (see additional figure containing CUT&RUN-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 21 Expand Image
    Immunoprecipitation of mineralocorticoid receptor protein from MSTO-211 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Mineralocorticoid Receptor (E9W1M) Rabbit mAb. Western blot analysis was performed using Mineralocorticoid Receptor (E9W1M) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 22 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and 5 μl of Estrogen Receptor α (D8H8) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 21 (upper), including TFF1/pS2 (lower), a known target gene of Estrogen Receptor α (see additional figure containing ChIP-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 23 Expand Image
    CUT&RUN was performed with MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 days, then treated with β-estradiol (10 nM) for 45 minutes and either Estrogen Receptor a (D8H8) Rabbit mAb or Estrogen Receptor α (D6R2W) Rabbit mAb #13258, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® #56795. The figures show binding across chromosome 21 (upper), including TFF1 gene (lower).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 24 Expand Image
    Western blot analysis of extracts from T-47D cells, grown for 48 hr in phenol red-free medium supplemented with 5% charcoal-stripped FBS and then treated with either a vehicle control (-) or promegestone (R5020, 100 nM, 16 hr; +), using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Prolonged treatment of PR-expressing cells with R5020 is known to induce PR downregulation and hyperphosphorylation, which is reflected by slower migration on SDS-PAGE.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 25 Expand Image
    CUT&RUN was performed with T-47D cells cultured in phenol red-free media supplemented with 5% charcoal-stripped FBS for 48 hr and then treated with promegestone (R5020, 10 nM, 1 hr) and Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 5 (upper), including SETD9 (lower).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 26 Expand Image
    Western blot analysis of extracts from control HeLa cells (lane 1), or Glucocorticoid Receptor knockout HeLa cells (lane 2) using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb #12041 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal in Glucocorticoid Receptor-knockout HeLa cells confirms specificity of the antibody for Glucocorticoid Receptor.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Androgen Receptor (D6F11) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 28 Expand Image
    CUT&RUN was performed with LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and either Androgen Receptor (D6F11) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human KLK2 Intron 1 Primers #62086 and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 29 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and either Estrogen Receptor α (D8H8) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 30 Expand Image
    CUT&RUN was performed with MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 days, then treated with β-estradiol (10 nM) for 45 minutes and either Estrogen Receptor a (D8H8) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human PS2 Promoter Primers #9702 and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human infiltrating ductal breast carcinoma using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 32 Expand Image
    CUT&RUN was performed with T-47D cells cultured in phenol red-free media supplemented with 5% charcoal-stripped FBS for 48 hr and then treated with promegestone (R5020, 10 nM, 1 hr) and either Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human TBL2 promoter primers and human OCM2 exon 1 primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 33 Expand Image
    Immunoprecipitation of glucocorticoid receptor from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 34 Expand Image
    Confocal immunofluorescent analysis of LNCaP (positive, left) and DU145 (negative, right) cells using Androgen Receptor (D6F11) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 35 Expand Image
    Immunohistochemical analysis of paraffin-embedded cell pellets, T-47D (high PR, left), MCF-7 (low PR, middle) and MDA-MB-231 (PR negative, right), using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 37 Expand Image
    Flow cytometric analysis of DU-145 cells (blue) and LNCaP cells (green) using Androgen Receptor (D6F11) XP® Rabbit mAb (solid lines) or a concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 39 Expand Image
    Confocal immunofluorescent analysis of T-47D (PR positive, left) and MDA-MB-231 (PR negative, right) cells using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse stomach using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 41 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and Androgen Receptor (D6F11) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across KLK2, a known target gene of Androgen Receptor (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 42 Expand Image
    Flow cytometric analysis of MDA-MB-231 cells (blue, negative) and T47D cells (green, positive) using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 43 Expand Image
    Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or dexamethasone-treated (right), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 44 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and Androgen Receptor (D6F11) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 19 (upper), including KLK2 (lower), a known target gene of Androgen Receptor (see additional figure containing ChIP-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 45 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from T-47D cells cultured in phenol red-free media supplemented with 5% charcoal-stripped FBS for 48 hr and then treated with promegestone (R5020, 10 nM, 1 hr) and Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across FKBP5/FKBP51, a known target gene of Progesterone Receptor A/B (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 47 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and either Androgen Receptor (D6F11) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human KLK2 Intron 1 Primers #62086, SimpleChIP® Human KLK3 Promoter Primers #32784, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 48 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from T-47D cells cultured in phenol red-free media supplemented with 5% charcoal-stripped FBS for 48 hr and then promegestone-treated (R5020, 10 nM, 1 hr) and Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 6 (upper), including FKBP5/ FKBP51 (lower), a known target gene of Progesterone Receptor A/B (see additional figure containing ChIP-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 49 Expand Image
    T-47D cells were cultured in phenol red-free media supplemented with 5% charcoal-stripped FBS for 48 hr and then either untreated (left panel) or promegestone-treated (R5020, 10 nM, 1 hr; right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin cells and Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human FKBP51 Intron 5 Primers #7859, human E2F-1 proximal enhancer site #1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 50 Expand Image
    Confocal immunofluorescent analysis of HeLa cells (left, positive) or HT-29 cells (right, negative) using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb #12041 (green), DyLight™ 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 51 Expand Image
    Flow cytometric analysis of HT-29 cells (blue, negative) and HeLa cells (green, positive) using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 52 Expand Image
    Human whole blood was fixed, lysed, and permeabilized as per the Cell Signaling Technology Flow Cytometry (Alternate) Protocol, and stained with CD3-PE, CD19-APC, and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb. B cell (green) and T cell (blue) population gates (left) were applied to a histogram depicting the mean fluorescence intensity of glucocorticoid receptor, compared to a nonspecific negative control antibody (red; right). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 53 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells cultured in media with 5% charcoal-stripped FBS for 3 d and then treated with 100 nM dexamethasone for 1 hr and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across SLC19A2, a known target gene of GR (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    Steroid Hormone Receptor Antibody Sampler Kit: Image 54 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells cultured in media with 5% charcoal-stripped FBS for 3 d and then treated with 100 nM dexamethasone for 1 hr and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 1 (upper), including SLC19A2 (lower), a known target gene of GR (see additional figure containing ChIP-qPCR data).
    Steroid Hormone Receptor Antibody Sampler Kit: Image 55 Expand Image
    A549 cells were cultured in media with 5% charcoal-stripped FBS for 3 d and then either untreated (left panel) or dexamethasone-treated (100 nM, 1 hr; right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human SLC19A2 Promoter Primers #7681, human MT2A promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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