LXR alpha/NR1H3 Antibody

Western Blot: LXR alpha/NR1H3 Antibody [NB300-612] - LXRs are expressed in peripheral blood cells. LXR alpha/NR1H3 protein levels in protein extracts from PBMCs from these same donors were detected by Western blotting using rabbit anti-human LXR alpha/NR1H3 polyclonal antisera. Image collected and cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-6-59), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: LXR alpha/NR1H3 Antibody [NB300-612] - Analysis of LXR alpha (green) showing staining in the nucleus of Hela cells (right) compared to a negative control without primary antibody (left).

Flow Cytometry: LXR alpha/NR1H3 Antibody [NB300-612] - analysis of LXR alpha was done on HeLa. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton X-100 for 20 minutes, and blocked with 2.5% BSA for 30 minutes at room temperature. Cells were labeled with LXR alpha Rabbit Polyclonal Antibody (PA1330, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Western Blot: LXR alpha/NR1H3 Antibody [NB300-612] - analysis was performed on nuclear enriched extracts (30 ug lysate) of MCF7 (Lane 1), THP-1 (Lane 2), A549 (Lane 3), SH-SY5Y (Lane 4), HL-60 (Lane 5) and KARPAS 299 (Lane 6). The blot was probed with Rabbit Anti-LXR alpha Polyclonal Antibody (2ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (0.25 ug/ml, 1:4000 dilution). A 50 kDa band corresponding to LXR alpha was observed across the cell lines tested. Known quantity of protein samples was electrophoresed using 4-12 % Bis-Tris gel, Electrophoresis System and Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with Blot 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using ECL Chemiluminescent Substrate Reagent Kit.

Immunocytochemistry/Immunofluorescence: LXR alpha/NR1H3 Antibody [NB300-612] - Analysis of LXR alpha (green) showing staining in the nucleus of MCF-7 cells (right) compared to a negative control without primary antibody (left).

LXRs are expressed in peripheral blood cells. (A) RNA from peripheral blood mononuclear cells obtained from normal human donors was assayed for LXR alpha and LXR beta transcript levels using qPCR. Expression values were normalized to GAPDH levels, represented as the mean +/- SEM. (B) LXR alpha protein levels in protein extracts from PBMCs from these same donors were detected by Western blotting using rabbit anti-human LXR alpha polyclonal antisera.

• Reactivity: Hu, Mu
• Applications: WB, Flow, GS, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

LXR alpha/NR1H3 Antibody Summary

• Immunogen: Synthetic peptide corresponding to residues C D(318) F S Y N R E D F A K A(329) of human LXR alpha.
• Specificity: Detects recombinant human LXR alpha. This does not detect recombinant human LXR beta.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: NR1H3
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 3-5ug/10^6 cells
  • Gel Super Shift Assays 1:1 - 1:100
  • Immunocytochemistry/ Immunofluorescence 1:50-1:500
  • Immunohistochemistry 1:10-1:500
  • Western Blot 1:300
• Application Notes: WB: Detects an approx. 64 kDa protein representing recombinant human LXR alpha.
  IHC usage was reported in the scientific literature (PMID: 17393442).
  Gel Shift Assays usage was reported in the scientific literature (PMID: 12470667).

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 17393442).

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS with 1 mg/ml BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for LXR alpha/NR1H3 Antibody

• alpha
• Liver X receptor alpha
• liver X receptor-alpha
• LXR alpha
• LXRA
• NR1H3
• Nuclear receptor subfamily 1 group H member 3
• nuclear receptor subfamily 1, group H, member 3
• oxysterols receptor LXR-alpha
• RLD-1

Background

Liver X Receptors (LXRs) are oxysterol activated nuclear receptors which are involved in the regulation of cholesterol metabolism and bile acids. Two known isoforms, alpha and beta, are differentially expressed. The alpha isoform is predominantly expressed in liver, whereas the beta isoform expression has been shown to be ubiquitous.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for LXR alpha/NR1H3 Antibody (NB300-612)(10)

Publications using NB300-612

• Palma GBH, Kaur M miRNA-128 and miRNA-223 regulate cholesterol-mediated drug resistance in breast cancer IUBMB life 2023-04-18 [PMID: 37070323] (WB, Human)
• Rudalska R, Harbig J, Snaebjornsson M et al. LXR alpha activation and Raf inhibition trigger lethal lipotoxicity in liver cancer Nature Cancer 2021-02-01 [PMID: 35122079]
• (WB, Mouse)
• Shen C, Jiang J, Huang C, Zhu W. Gypenoside LVI attenuates foam cell formation by promoting cholesterol export and inhibiting inflammation response Oncotarget 2018-10-05
• Bruschi FV, Claudel T, Tardelli M et al. PNPLA3 I148M Variant Impairs Liver X Receptor Signaling and Cholesterol Homeostasis in Human Hepatic Stellate Cells Hepatol Commun 2019-09-01 [PMID: 31497741] (WB, Human)
• Abuirqeba SA The Role of Liver X Receptor in Prostate Cancer Metabolism Thesis 1905-07-13
• Poomthavorn P, Wong SHX, Higgins S et al. Activation of a prometastatic gene expression program in hypoxic neuroblastoma cells. Endocr Relat Cancer;16(3):991-1004. 2009-01-01 [PMID: 19423615]
• Dang H, Liu Y, Pang W et al. Suppression of 2,3-Oxidosqualene Cyclase by High Fat Diet Contributes to Liver X Receptor-{alpha}-mediated Improvement of Hepatic Lipid Profile. J Biol Chem;284(10):6218-6226. 2009-01-01 [PMID: 19119143]
• DiBlasio-Smith EA, Arai M, Quinet EM et al. Discovery and implementation of transcriptional biomarkers of synthetic LXR agonists in peripheral blood cells (BioMed Central). J Transl Med. 2008-01-01 [PMID: 18925943]
• Zhang, C et al. NO-1886 suppressing atherosclerosis in high-fat-high-sucrose/high-cholesterol fed Bama minipigs is related to upregulating ATP-binding cassette transporter A1. J Lipid Res. 2006-01-01 [PMID: 16807312]

Bioinformatics

• Gene Symbol: NR1H3