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8595
Nuclear Receptor Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Nuclear Receptor Antibody Sampler Kit #8595

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Nuclear Receptor Antibody Sampler Kit: Image 1
Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells (differentiated 6 days into adipocytes) using PPARγ (C26H12) Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 2
Western blot analysis of extracts from various cell lines using RXRα (D6H10) Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 3
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a construct 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 (D8H2) XP® Rabbit mAb (upper) and DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368 (lower).
Nuclear Receptor Antibody Sampler Kit: Image 4
Confocal immunofluorescent analysis of HeLa cells (left, positive) or HT-29 cells (right, negative) using Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660 (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
Nuclear Receptor Antibody Sampler Kit: Image 5
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).
Nuclear Receptor Antibody Sampler Kit: Image 6
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 Library was prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across KLK2, a known target gene of Androgen Receptor (see additional figure containing CUT&RUN-qPCR data).
Nuclear Receptor Antibody Sampler Kit: Image 7
Western blot analysis of extracts from various cell lines using RARα (E6Z6K) Rabbit mAb. The NB-4 cell line contains the PML-RARα fusion protein.
Nuclear Receptor Antibody Sampler Kit: Image 8
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.
Nuclear Receptor Antibody Sampler Kit: Image 9
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 Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across TFF1/pS2, a known target gene of Estrogen Receptor α (see additional figure containing ChIP-qPCR data).
Nuclear Receptor Antibody Sampler Kit: Image 10
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).
Nuclear Receptor Antibody Sampler Kit: Image 11
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 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across TFF1 gene.
Nuclear Receptor Antibody Sampler Kit: Image 12
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).
Nuclear Receptor Antibody Sampler Kit: Image 13
Western blot analysis of extracts from various cell lines using RARγ1 (D3A4) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 14
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with Myc/DDK-tagged cDNA expression constructs encoding full-length human RXRα (hRXRα; +), RXRβ (hRXRβ; +), or RXRγ (hRXRγ; +), using RXRα (D6H10) Rabbit mAb (upper) and DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368 (lower).
Nuclear Receptor Antibody Sampler Kit: Image 15
Western blot analysis of extracts from various cell lines using Glucocorticoid Receptor (D8H2) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded LNCaP (AR+, left) and DU145 (AR-, right) using Androgen Receptor (D6F11) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 17
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 Library was prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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).
Nuclear Receptor Antibody Sampler Kit: Image 18
Immunoprecipitation of RARα and PML-RARα from NB-4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is RARα (E6Z6K) Rabbit mAb. Western blot analysis was performed with RARα (E6Z6K) Rabbit mAb. The NB-4 cell line contains the PML-RARα fusion. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Nuclear Receptor Antibody Sampler Kit: Image 19
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 Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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).
Nuclear Receptor Antibody Sampler Kit: Image 20
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 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figures show binding across chromosome 21 (upper), including TFF1 gene (lower).
Nuclear Receptor Antibody Sampler Kit: Image 21
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.
Nuclear Receptor Antibody Sampler Kit: Image 22
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length human RARγ1 (hRARγ1-Myc/DDK, +), using RARγ1 (D3A4) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 23
Immunohistochemical analysis of 3T3-L1 cells, undifferentiated (left) or differentiated (right) , using PPARγ (C26H12) Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 24
Chromatin immunoprecipitations were performed with cross-linked chromatin from LS-180 cells treated with Vitamin D (10nM) for 3 hours and either RXRα (D6H10) 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 c-Fos Upstream Primers #25661, human UCA1 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.
Nuclear Receptor Antibody Sampler Kit: Image 25
Western blot analysis of extracts from control HeLa cells (lane 1), or Glucocorticoid Receptor knockout HeLa cells (lane 2) using Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the Glucocorticoid Receptor-knockout HeLa cells confirms specificity of the antibody for Glucocorticoid Receptor.
Nuclear Receptor Antibody Sampler Kit: Image 26
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Androgen Receptor (D6F11) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 27
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.
Nuclear Receptor Antibody Sampler Kit: Image 28
Chromatin immunoprecipitations were performed with cross-linked chromatin from NB-4 cells and either RARα (E6Z6K) Rabbit mAb #62294 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 RUNX1 Exon 2 Primers #67254, human GFI promoter primers, human GFI exon 4 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.
Nuclear Receptor Antibody Sampler Kit: Image 29
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.
Nuclear Receptor Antibody Sampler Kit: Image 30
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.
Nuclear Receptor Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded human infiltrating ductal breast carcinoma using Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded cell pellets, HaCaT (positive, left) and Hep3B (negative, right), using RARγ1 (D3A4) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded mouse brown fat using PPARγ (C26H12) Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 34
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).
Nuclear Receptor Antibody Sampler Kit: Image 35
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.
Nuclear Receptor Antibody Sampler Kit: Image 36
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using RARγ1 (D3A4) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 37
Confocal immunofluorescent analysis of 3T3-L1 cells using PPARγ (C26H12A8) Rabbit mAb (red) showing nuclear localization in differentiated cells. Lipid droplets have been labeled with BODIPY 493/503 (green). Blue pseudocolor = DRAQ5 (fluorescent DNA dye).
Nuclear Receptor Antibody Sampler Kit: Image 38
Flow cytometric analysis of HT-29 cells (blue, negative) and HeLa cells (green, positive) using Glucocorticoid Receptor (D8H2) 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.
Nuclear Receptor Antibody Sampler Kit: Image 39
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.
Nuclear Receptor Antibody Sampler Kit: Image 40
Immunohistochemical analysis of paraffin-embedded human skin using RARγ1 (D3A4) XP® Rabbit mAb.
Nuclear Receptor Antibody Sampler Kit: Image 41
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).
Nuclear Receptor Antibody Sampler Kit: Image 42
Confocal immunofluorescent analysis of HaCaT cells (positive, left) and Hep3B cells (negative, right) using RARγ1 (D3A4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Nuclear Receptor Antibody Sampler Kit: Image 43
Chromatin immunoprecipitations were performed with cross-linked chromatin from HDLM-2 cells and either PPARγ (C26H12) 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 FOXN3 Intron 3 Primers #95568, human STON2 intron 4 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.
Nuclear Receptor Antibody Sampler Kit: Image 44
Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells cultured in media with 5% charcoal-stripped FBS for 3 days and then dexamethasone-treated (100 nM, 1 hr) and Glucocorticoid Receptor (D8H2) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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 data sheet.
Nuclear Receptor Antibody Sampler Kit: Image 45
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 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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 data sheet.
Nuclear Receptor Antibody Sampler Kit: Image 46
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.
Nuclear Receptor Antibody Sampler Kit: Image 47
Flow cytometric analysis of T-47D cells using RARγ1 (D3A4) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Nuclear Receptor Antibody Sampler Kit: Image 48
Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells cultured in media with 5% charcoal-stripped FBS for 3 days and then dexamethasone-treated (100 nM, 1 hr) and Glucocorticoid Receptor (D8H2) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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).
Nuclear Receptor Antibody Sampler Kit: Image 49
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 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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).
Nuclear Receptor Antibody Sampler Kit: Image 50
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 Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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 data sheet.
Nuclear Receptor Antibody Sampler Kit: Image 51
A549 cells were cultured in media with 5% charcoal-stripped FBS for 3 days and then either untreated (left panel) or dexamethasone-treated (100 nM, 1 hr; right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin and Glucocorticoid Receptor (D8H2) 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.
Nuclear Receptor Antibody Sampler Kit: Image 52
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.
Nuclear Receptor Antibody Sampler Kit: Image 53
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 Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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).
Nuclear Receptor Antibody Sampler Kit: Image 54
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.
To Purchase # 8595
Cat. # Size Qty. Price
8595T
1 Kit  (8 x 20 microliters) $ 592

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
RARα (E6Z6K) Rabbit mAb 62294 20 µl
  • WB
  • IP
  • ChIP
H M Mk 60 Rabbit IgG
RARγ1 (D3A4) XP® Rabbit mAb 8965 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M 58 Rabbit IgG
RXRα (D6H10) Rabbit mAb 3085 20 µl
  • WB
  • IP
  • ChIP
H M R 53 Rabbit IgG
Glucocorticoid Receptor (D8H2) XP® Rabbit mAb 3660 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M R Mk 80, 91, 94 Rabbit IgG
Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb 8757 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H 90 (PR-A), 118 (PR-B) Rabbit IgG
Androgen Receptor (D6F11) XP® Rabbit mAb 5153 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
  • C&R
H 110 Rabbit IgG
Estrogen Receptor α (D8H8) Rabbit mAb 8644 20 µl
  • WB
  • IP
  • ChIP
  • C&R
H 66 Rabbit IgG
PPARγ (C26H12) Rabbit mAb 2435 20 µl
  • WB
  • IHC
  • IF
  • ChIP
H M 53, 57 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

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 two 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. RARα (E6Z6K) Rabbit mAb weakly detects RARγ when it is overexpressed.

Source / Purification

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, residues near the amino terminus of human RXRα protein, or residues surrounding Leu220 of human RARα protein.

Background

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).
  1. Li, J. and Al-Azzawi, F. (2009) Maturitas 63, 142-8.
  2. Avila, D.M. et al. (2010) 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. Yamamoto, K.R. (1985) Annu Rev Genet 19, 209-52.
  7. Tontonoz, P. et al. (1995) Curr Opin Genet Dev 5, 571-6.
  8. Rosen, E.D. et al. (1999) Mol Cell 4, 611-7.
  9. Murphy, G.J. and Holder, J.C. (2000) Trends Pharmacol Sci 21, 469-74.
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