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8589
Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit #8589

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Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 1

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.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 2

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.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 3

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 RXRα, RXRβ, and RXRγ, using RXRβ Antibody (upper) or DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368 (lower).

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 4

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.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 5

Western blot analysis of extracts from 293T cells, either mock transfected or transfected with human RXRα, RXRβ, or RXRγ DYKDDDK-tagged constructs, using RXRγ Antibody (upper) and DYKDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368 (lower).

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 6

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.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 7

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.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 8

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).

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 9

Western blot analysis of extracts from various cell lines using RXRβ Antibody.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 10

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).

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 11

Western blot analysis of extracts from various tissues using RXRγ Antibody.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 12

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.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 13

Western blot analysis of extracts from various cell lines using RXRα (D6H10) Rabbit mAb.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 14

Immunohistochemical analysis of paraffin-embedded cell pellets, HaCaT (positive, left) and Hep3B (negative, right), using RARγ1 (D3A4) XP® Rabbit mAb.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 15

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using RARγ1 (D3A4) XP® Rabbit mAb.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 16

Immunohistochemical analysis of paraffin-embedded human skin using RARγ1 (D3A4) XP® Rabbit mAb.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 17

Western blot analysis of extracts from various cell lines using RARγ1 (D3A4) XP® Rabbit mAb.

Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit: Image 18

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.

To Purchase # 8589T
Product # Size Price
8589T
1 Kit  (5 x 20 µl) $ 391

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
RARα (E6Z6K) Rabbit mAb 62294 20 µl
  • WB
  • IP
  • ChIP
H M Mk 60 Rabbit IgG
RXRα (D6H10) Rabbit mAb 3085 20 µl
  • WB
  • IP
  • ChIP
H M R 53 Rabbit IgG
RXRβ Antibody 8715 20 µl
  • WB
  • IP
H M 70-72 Rabbit 
RARγ1 (D3A4) XP® Rabbit mAb 8965 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M 58 Rabbit IgG
RXRγ Antibody 5629 20 µl
  • WB
  • IP
H M 55 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit provides an economical means to investigate the expression of various subtypes of retinoic acid and retinoid X receptors. The kit contains enough primary antibody to perform two western blot experiments per primary.

Specificity / Sensitivity

Each antibody in the Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit recognizes endogenous levels of total respective protein. The antibodies do not cross react with other subtypes of retinoic acid or retinoic X receptors, with the exception of RARα (E6Z6K) Rabbit mAb, which may weakly detect RARγ when it is overexpressed.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human RARγ1 protein, the amino terminus of human RXRα protein, or residues surrounding Leu220 of human RARα protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human RXRβ protein, or residues near the amino terminus of human RXRγ protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Nuclear retinoic acid (RA) 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 (1). 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 (2,3). Consequently, RARs play critical roles in a variety of biological processes, including development, reproduction, immunity, and organogenesis (4-6). RAR mutations, fusion proteins, altered expression levels, or aberrant post-translational modifications result in multiple diseases due to altered RAR function and disruption of homeostasis.

In contrast to the ubiquitously expressed RARα subtype, RARγ displays a complex tissue-specific expression pattern (7). The hematopoietic system expresses significant levels of RARγ, and a recent study identified a role for RARγ in hematopoietic stem cell maintenance (8). RARγ is the predominant subtype in human and mouse epidermis, representing 90% of the RARs in this tissue (9-11). Given the high level of RARγ expression in the skin, it has been suggested that this nuclear receptor participates in a transcriptional program that governs maintenance and differentiation of normal epidermis and skin appendages. The transcriptional activity of RARγ is under stringent control, in part, through retinoic acid-induced phosphorylation and proteasomal degradation (12).

The human retinoid X receptors (RXRs) 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 (13). Since RXRs heterodimerize with multiple nuclear hormone receptors, they play a central role in transcriptional control of numerous hormonal signaling pathways by binding to cis-acting response elements in the promoter/enhancer region of target genes (14).

  1. Rochette-Egly, C. and Germain, P. (2009) Nucl Recept Signal 7, e005.
  2. Delacroix, L. et al. (2010) Mol Cell Biol 30, 231-44.
  3. Eifert, C. et al. (2006) Mol Reprod Dev 73, 796-824.
  4. Mark, M. et al. (2006) Annu Rev Pharmacol Toxicol 46, 451-80.
  5. Niederreither, K. and Dollé, P. (2008) Nat Rev Genet 9, 541-53.
  6. Mark, M. et al. (2009) Nucl Recept Signal 7, e002.
  7. Dollé, P. (2009) Nucl Recept Signal 7, e006.
  8. Purton, L.E. et al. (2006) J Exp Med 203, 1283-93.
  9. Fisher, G.J. et al. (1994) J Biol Chem 269, 20629-35.
  10. Zelent, A. et al. (1989) Nature 339, 714-7.
  11. Elder, J.T. et al. (1991) J Invest Dermatol 96, 425-33.
  12. Giannì, M. et al. (2002) EMBO J 21, 3760-9.
  13. Gronemeyer, H. et al. (2004) Nat Rev Drug Discov 3, 950-64.
  14. Mangelsdorf, D.J. et al. (1992) Genes Dev 6, 329-44.

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.
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