#8589 Retinoic Acid and Retinoid X Receptors Antibody Sampler Kit
|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||H, M, R||53||Rabbit IgG|
|RXRβ Antibody #8715||20 µl||WB, IP||H, M||R, Mk, B, Dg, Pg||70-72||Rabbit|
|RARγ1 (D3A4) XP® Rabbit mAb #8965||20 µl||WB, IP, IHC-P, IF-IC, F||H, M||R, Hm, B, Dg||58||Rabbit IgG|
|RXRγ Antibody #5629||20 µl||WB, IP||H, M||R, Mk, B, Dg||55||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||WB||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, Mk=Monkey, R=Rat
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.
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).
Western blot analysis of extracts from various cell lines using RARγ1 (D3A4) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using RXRα (D6H10) Rabbit mAb.
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.
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).
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.
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).
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.
Immunohistochemical analysis of paraffin-embedded cell pellets, HaCaT (positive, left) and Hep3B (negative, right), using RARγ1 (D3A4) XP® Rabbit mAb.
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.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using RARγ1 (D3A4) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human skin using RARγ1 (D3A4) XP® Rabbit mAb.
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 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).
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Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Anti-FLAG is a registered trademark of Sigma-Aldrich Biotechnology.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.