LDLR Antibody [Unconjugated]

LDL R was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human LDL R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2148) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

LDL R was detected in formalin fixed paraffin-embedded sections of human liver using Goat Anti-Human LDL R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2148) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

APLP2 and LDLR interactions with PCSK9 and their regulation of PCSK9 function.(A and B) Western blot showing APLP2, PCSK9, or Transferrin receptor (TFNR) levels in input fraction (I), IC or J16 immunoprecipitated samples (IP Ab.) in the absence or presence of 5F6 Fab or 12E3 Fab, as indicated. (B) Quantification of (A); shown as average APLP2 normalized to PCSK9 of 3 independent experiments with SEM. (C and D) J16 coIPs of PCSK9 from Neg or LDLR siRNA treated HepG2 cells with IC control, as indicated. (D) Quantification of (C); shown as average APLP2 normalized to PCSK9 from 3 independent experiments with SEM. (E, F, and G) Western blot of LDLR, APOER2, or TFNR in siRNA treated cells following treatment with PCSK9 at 0, 20, 50, or 100 μg/ml. (F) LDLR levels from (E) quantified as percent LDLR degradation of untreated cells and normalized to Neg siRNA samples. Shown as average with SEM from 4 independent experiments. (G) Same as F, but measuring APOER2 levels. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25905719), licensed under a CC-BY license. Not internally tested by R&D Systems.

APLP2 and LDLR interactions with PCSK9 and their regulation of PCSK9 function.(A and B) Western blot showing APLP2, PCSK9, or Transferrin receptor (TFNR) levels in input fraction (I), IC or J16 immunoprecipitated samples (IP Ab.) in the absence or presence of 5F6 Fab or 12E3 Fab, as indicated. (B) Quantification of (A); shown as average APLP2 normalized to PCSK9 of 3 independent experiments with SEM. (C and D) J16 coIPs of PCSK9 from Neg or LDLR siRNA treated HepG2 cells with IC control, as indicated. (D) Quantification of (C); shown as average APLP2 normalized to PCSK9 from 3 independent experiments with SEM. (E, F, and G) Western blot of LDLR, APOER2, or TFNR in siRNA treated cells following treatment with PCSK9 at 0, 20, 50, or 100 μg/ml. (F) LDLR levels from (E) quantified as percent LDLR degradation of untreated cells and normalized to Neg siRNA samples. Shown as average with SEM from 4 independent experiments. (G) Same as F, but measuring APOER2 levels. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25905719), licensed under a CC-BY license. Not internally tested by R&D Systems.

Assessment of PCSK9 and LDLR expression in human LEC. (A) PCSK9 expression was measured by immunoblotting in either human LEC, HepG2 or HEK 293T cell lysates. (B) ELISA was used to measure PCSK9 levels in the cell culture supernatant of Huh7 (grey), HEK 293T (white) and human LEC (black). (C) Expression of LDLR was detected in protein lysates by immunoblotting of human LEC. Huh7 cells were used as a positive control and HEK 293T cells treated with siLDLR were used as a negative control. LDLR protein expression on human LEC was measured by (D) flow cytometry after extracellular staining of LEC (black line) and Huh7 (grey line) and by (E) immunofluorescence (Blue, DAPI; red, cholera toxin; green, anti-LDLR; yellow, colocalized voxels; scale bar, 20 µM). (F) Scatterplot of red and green pixel intensities of cholera toxin (red) and anti-LDLR (green) in human LEC. n = 4-9. Statistics: ****p < 0.0001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35154499), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

LDLR Antibody [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human LDL R
  Asp193-Arg788
  Accession # P01130
• Specificity: Detects human LDL R in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant mouse LDL R is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: LDLR
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Blockade of Receptor-ligand Interaction
  • Immunocytochemistry 1-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for LDLR Antibody [Unconjugated]

• FH
• FHC
• LDL R
• LDL receptor
• LDLCQ2
• LDLR
• low density lipoprotein receptor
• low-density lipoprotein receptor class A domain-containing protein 3
• low-density lipoprotein receptor

Background

The low density lipoprotein receptor (LDL R) is the founding member of the LDL R family of scavenger receptors. This family contains transmembrane molecules that are characterized by the presence of EGF repeats, complement-like repeats, and YWTD motifs that form beta -propellers. Although members of the family were originally thought to be endocytic receptors, it is now clear that some members interact with adjacent cell-surface molecules, expanding their range of activities. Human LDL R is synthesized as an 860 amino acid (aa) precursor that contains a 21 aa signal sequence, a 767 aa extracellular region, a 22 aa transmembrane segment and a 50 aa cytoplasmic tail. The extracellular region is complex. It consists of seven N-terminal complement-like cysteine-rich repeats that bind ligand. Cysteine residues in this region participate in intrachain disulfide bonds. This region is followed by three EGF-like repeats with a beta -propeller YWTD containing motif. The EGF-like repeats are responsible for ligand bonding and dissociation. Finally, there is a 50 aa membrane proximal Ser/Thr-rich region that serves as a carbohydrate attachment point. There is extensive O-linked and modest N-linked glycosylation. Thus the receptor’s predicted molecular weight of 93 kDa is increased to a native molecular weight of 120 - 160 kDa. Within the 50 aa cytoplasmic tail, there is an NPXY motif that links the receptor to clathrin pits. The extracellular region of human LDL R is 51% aa identical to the extracellular region of human VLDL R, and 79% aa identical to the extracellular region of mouse LDL R. LDL R is constitutively expressed and binds apoB of LDL and apoE of VLDL. It is responsible for clearing 70% of plasma LDL in liver. Mutations in the LDL R gene cause the autosomal dominant disorder, familial hypercholesterolemia.

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 LDLR Antibody (AF2148)(39)

Publications using AF2148

• Ritter, M;Canus, L;Gautam, A;Vallet, T;Zhong, L;Lalande, A;Boson, B;Gandhi, A;Bodoirat, S;Burlaud-Gaillard, J;Freitas, N;Roingeard, P;Barr, JN;Lotteau, V;Legros, V;Mathieu, C;Cosset, FL;Denolly, S; The low-density lipoprotein receptor and apolipoprotein E associated with CCHFV particles mediate CCHFV entry into cells Nature communications 2024-05-28 [PMID: 38806525] (Flow Cytometry, Western Blot, Neutralization, Immunocytochemistry, Human, Primate - Cercopithecus aethiops (African Green Monkey))
• Mikaeeli, S;Ben Djoudi Ouadda, A;Evagelidis, A;Essalmani, R;Ramos, OHP;Fruchart-Gaillard, C;Seidah, NG; Insights into PCSK9-LDLR Regulation and Trafficking via the Differential Functions of MHC-I Proteins HFE and HLA-C Cells 2024-05-17 [PMID: 38786080] (Western Blot, Immunocytochemistry, Human)
• Magalhães, A;Cesário, V;Coutinho, D;Matias, I;Domingues, G;Pinheiro, C;Serafim, T;Dias, S; A high-cholesterol diet promotes the intravasation of breast tumor cells through an LDL-LDLR axis Scientific reports 2024-04-24 [PMID: 38658568] (Functional Assay, Human, Mouse)
• Zakharova, IS;Shevchenko, AI;Arssan, MA;Sleptcov, AA;Nazarenko, MS;Zarubin, AA;Zheltysheva, NV;Shevchenko, VA;Tmoyan, NA;Saaya, SB;Ezhov, MV;Kukharchuk, VV;Parfyonova, YV;Zakian, SM; iPSC-Derived Endothelial Cells Reveal LDLR Dysfunction and Dysregulated Gene Expression Profiles in Familial Hypercholesterolemia International journal of molecular sciences 2024-01-05 [PMID: 38255763] (Western Blot, Human)
• Vadon-Le Goff, S;Tessier, A;Napoli, M;Dieryckx, C;Bauer, J;Dussoyer, M;Lagoutte, P;Peyronnel, C;Essayan, L;Kleiser, S;Tueni, N;Bettler, E;Mariano, N;Errazuriz-Cerda, E;Fruchart Gaillard, C;Ruggiero, F;Becker-Pauly, C;Allain, JM;Bruckner-Tuderman, L;Nyström, A;Moali, C; Identification of PCPE-2 as the endogenous specific inhibitor of human BMP-1/tolloid-like proteinases Nature communications 2023-12-04 [PMID: 38049428] (Western Blot, Mouse, Human)
• Alexandra Fowler, Koen K. A. Van Rompay, Maureen Sampson, Javier Leo, Jennifer K. Watanabe, Jodie L. Usachenko et al. A virus-like particle-based bivalent PCSK9 vaccine lowers LDL-cholesterol levels in non-human primates NPJ Vaccines 9/28/2023 [PMID: 37770440]
• Fowler, A;Van Rompay, KKA;Sampson, M;Leo, J;Watanabe, JK;Usachenko, JL;Immareddy, R;Lovato, DM;Schiller, JT;Remaley, AT;Chackerian, B; A Virus-like particle-based bivalent PCSK9 vaccine lowers LDL-cholesterol levels in Non-Human Primates bioRxiv : the preprint server for biology 2023-05-15 [PMID: 37292981] (Western Blot, Primate - Macaca fascicularis (Cynomolgus Macaque))
• Suzanne E. Engelen, Francesca A. Ververs, Angela Markovska, B. Christoffer Lagerholm, Jordan M. Kraaijenhof, Laura I.E. Yousif et al. Lipoproteins act as vehicles for lipid antigen delivery and activation of invariant natural killer T cells JCI Insight 5/8/2023 [PMID: 36976644]
• Manel Essaidi-Laziosi, Léna Royston, Bernadett Boda, Francisco Javier Pérez-Rodriguez, Isabelle Piuz, Nicolas Hulo et al. Altered cell function and increased replication of rhinoviruses and EV-D68 in airway epithelia of asthma patients Frontiers in Microbiology 3/1/2023 [PMID: 36937308]
• V Jain, SL Harper, AM Versace, D Fingerman, GS Brown, M Bhardwaj, MAS Crissey, AR Goldman, G Ruthel, Q Liu, A Zivkovic, H Stark, M Herlyn, PA Gimotty, DW Speicher, RK Amaravadi Targeting UGCG overcomes resistance to lysosomal autophagy inhibition Cancer Discovery, 2023-02-06;0(0):. 2023-02-06 [PMID: 36331284] (Neutralization, Western Blot, Human)
• Laurent Vachon, Ali Smaani, Nolwenn Tessier, Gabriel Jean, Annie Demers, Andreea Milasan et al. Downregulation of low-density lipoprotein receptor mRNA in lymphatic endothelial cells impairs lymphatic function through changes in intracellular lipids Theranostics 1/1/2022 [PMID: 35154499]
• Angélina Acier, Magali Godard, Fanny Gassiot, Pascal Finetti, Marion Rubis, Jonathan Nowak et al. LDL receptor-peptide conjugate as in vivo tool for specific targeting of pancreatic ductal adenocarcinoma Communications Biology 8/19/2021 [PMID: 34413441]
• JA Levine, C Oleaga, M Eren, AP Amaral, M Shang, E Lux, SS Khan, SJ Shah, Y Omura, N Pamir, J Hay, G Barish, T Miyata, H Tavori, S Fazio, DE Vaughan Role of PAI-1 in hepatic steatosis and dyslipidemia Scientific Reports, 2021-01-11;11(1):430. 2021-01-11 [PMID: 33432099] (ICC, Human)
• Chang Zhang, Chanjuan Hao, Guanghou Shui, Wei Li BLOS1 mediates kinesin switch during endosomal recycling of LDL receptor eLife 11/12/2020 [PMID: 33179593]
• S Banerjee, RJ Andrew, CJ Duff, K Fisher, CD Jackson, CB Lawrence, N Maeda, DS Greenspan, KAB Kellett, NM Hooper Author Correction: Proteolysis of the low density lipoprotein receptor by bone morphogenetic protein-1 regulates cellular cholesterol uptake Sci Rep, 2020-02-11;10(1):2681. 2/11/2020 [PMID: 32042031]
• Sreemoti Banerjee, Robert J. Andrew, Christopher J. Duff, Kate Fisher, Carolyn D. Jackson, Catherine B. Lawrence et al. Proteolysis of the low density lipoprotein receptor by bone morphogenetic protein-1 regulates cellular cholesterol uptake Scientific Reports 8/6/2019 [PMID: 31388055]
• L Huang, KL Chambliss, X Gao, IS Yuhanna, E Behling-Ke, S Bergaya, M Ahmed, P Michaely, K Luby-Phelp, A Darehshour, L Xu, EA Fisher, WP Ge, C Mineo, PW Shaul SR-B1 drives endothelial cell LDL transcytosis via DOCK4 to promote atherosclerosis Nature, 2019-04-24;0(0):. 2019-04-24 [PMID: 31019307] (Western Blot, Human)
• V Sachan, R Lodge, K Mihara, J Hamelin, C Power, BB Gelman, MD Hollenberg, ÉA Cohen, NG Seidah HIV-induced neuroinflammation: impact of PAR1 and PAR2 processing by Furin Cell Death Differ., 2019-01-25;0(0):. 2019-01-25 [PMID: 30683917] (ICC, Human)
• N Volkmar, ML Thezenas, SM Louie, S Juszkiewic, DK Nomura, RS Hegde, BM Kessler, JC Christians The ER membrane protein complex promotes biogenesis of sterol-related enzymes maintaining cholesterol homeostasis J. Cell. Sci., 2019-01-16;132(2):. 2019-01-16 [PMID: 30578317] (Western Blot, Human)
• PM Grin, DJ Dwivedi, KM Chathely, BL Trigatti, A Prat, NG Seidah, PC Liaw, AE Fox-Robich Low-density lipoprotein (LDL)-dependent uptake of Gram-positive lipoteichoic acid and Gram-negative lipopolysaccharide occurs through LDL receptor Sci Rep, 2018-07-12;8(1):10496. 2018-07-12 [PMID: 30002483] (Functional Assay, Human)
• X Li, J Guo, N Liang, X Jiang, Y Song, S Ou, Y Hu, R Jiao, W Bai 6-Gingerol Regulates Hepatic Cholesterol Metabolism by Up-regulation of LDLR and Cholesterol Efflux-Related Genes in HepG2 Cells Front Pharmacol, 2018-02-27;9(0):159. 2018-02-27 [PMID: 29535632] (Western Blot, Human)
• XH Liang, H Sun, W Shen, S Wang, J Yao, MT Migawa, HH Bui, SS Damle, S Riney, MJ Graham, RM Crooke, ST Crooke Antisense oligonucleotides targeting translation inhibitory elements in 5&#039; UTRs can selectively increase protein levels Nucleic Acids Res., 2017-09-19;45(16):9528-9546. 2017-09-19 [PMID: 28934489] (Western Blot, Human)
• SA Felt, GN Droby, VZ Grdzelishv Ruxolitinib and polycation combination treatment overcomes multiple mechanisms of resistance of pancreatic cancer cells to oncolytic vesicular stomatitis virus J. Virol., 2017-07-27;0(0):. 2017-07-27 [PMID: 28566376] (Western Blot, Human)
• R Velázquez-, M Law, J Bukh, J Prentoe Applying antibody-sensitive hypervariable region 1-deleted hepatitis C virus to the study of escape pathways of neutralizing human monoclonal antibody AR5A PLoS Pathog, 2017-02-23;13(2):e1006214. 2017-02-23 [PMID: 28231271] (Neutralization, Human)
• Delia Susan-Resi A novel mechanism causing familial hypercholesterolemia: The proprotein convertase subtilisin/kexin type 9-resistant Arg410Ser LDL receptor mutation J. Biol. Chem, 2016-12-20;0(0):. 2016-12-20 [PMID: 27998977] (ICC, Western Blot, Human)
• Identification of a Chrysanthemic Ester as an Apolipoprotein E Inducer in Astrocytes PLoS ONE, 2016-09-06;11(9):e0162384. 2016-09-06 [PMID: 27598782] (Western Blot, Human)
• Emilienne Tudor Ngo Sock, Gaétan Mayer, Jean-Marc Lavoie Combined Effects of Rosuvastatin and Exercise on Gene Expression of Key Molecules Involved in Cholesterol Metabolism in Ovariectomized Rats PLOS ONE 7/21/2016 [PMID: 27442011]
• Elodie Weider Proprotein convertase subtilisin/Kexin type 9 (PCSK9) single domain antibodies are potent inhibitors of LDL receptor degradation J Biol Chem, 2016-06-08;0(0):. 2016-06-08 [PMID: 27284008] (Western Blot, Human)
• Steve Poirier, Hocine Ait Hamouda, Louis Villeneuve, Annie Demers, Gaétan Mayer Trafficking Dynamics of PCSK9-Induced LDLR Degradation: Focus on Human PCSK9 Mutations and C-Terminal Domain PLOS ONE 6/9/2016 [PMID: 27280970]
• Alirocumab, a Therapeutic Human Antibody to PCSK9, Does Not Affect CD81 Levels or Hepatitis C Virus Entry and Replication into Hepatocytes PLoS ONE, 2016-04-26;11(4):e0154498. 2016-04-26 [PMID: 27115873] (Western Blot, Human)
• Le Q, Blanchet M, Seidah N, Labonte P Plasma Membrane Tetraspanin CD81 Complexes with Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and Low Density Lipoprotein Receptor (LDLR), and Its Levels Are Reduced by PCSK9. J Biol Chem, 2015-07-20;290(38):23385-400. 2015-07-20 [PMID: 26195630] (IHC, Immunoprecipitation, Human)
• DeVay R, Yamamoto L, Shelton D, Liang H Common Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) Epitopes Mediate Multiple Routes for Internalization and Function. PLoS ONE, 2015-04-23;10(4):e0125127. 2015-04-23 [PMID: 25905719] (IHC, Western Blot, Human, Mouse)
• Klop B, van der Pol P, van Bruggen R, Wang Y, de Vries M, van Santen S, O'Flynn J, van de Geijn G, Njo T, Janssen H, de Man P, Jukema J, Rabelink T, Rensen P, van Kooten C, Cabezas M Differential complement activation pathways promote C3b deposition on native and acetylated LDL thereby inducing lipoprotein binding to the complement receptor 1. J Biol Chem, 2014-10-27;289(51):35421-30. 2014-10-27 [PMID: 25349208] (Blocking, Flow Cytometry, Hamster, Human)
• Christian K. Mathiesen, Tanja B. Jensen, Jannick Prentoe, Henrik Krarup, Alfredo Nicosia, Mansun Law et al. Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1–6 Virology 6/1/2014 [PMID: 24928051]
• Jannick Prentoe, Stéphanie B. N. Serre, Santseharay Ramirez, Alfredo Nicosia, Judith M. Gottwein, Jens Bukh Hypervariable Region 1 Deletion and Required Adaptive Envelope Mutations Confer Decreased Dependency on Scavenger Receptor Class B Type I and Low-Density Lipoprotein Receptor for Hepatitis C Virus Journal of Virology 2/1/2014 [PMID: 24257605]
• Fan , Jianjia, Shimizu , Yoko, Chan , Jeniffer, Wilkinson , Anna, Ito , Ayaka, Tontonoz , Peter, Dullaghan , Edie, Galea , Liisa A, Pfeifer , Tom, Wellington , Cheryl L Hormonal modulators of glial ABCA1 and apoE levels. J Lipid Res, 2013-09-02;54(11):3139-50. 2013-09-02 [PMID: 23999864] (Western Blot, Human)
• DeVay , Rachel M, Shelton , David L, Liang , Hong Characterization of proprotein convertase subtilisin/kexin type 9 (PCSK9) trafficking reveals a novel lysosomal targeting mechanism via amyloid precursor-like protein 2 (APLP2). J Biol Chem, 2013-02-19;288(15):10805-18. 2013-02-19 [PMID: 23430252] (ICC, Western Blot, Human)
• Rousselet E, Benjannet S, Hamelin J, Canuel M, Seidah NG The proprotein convertase PC7: unique zymogen activation and trafficking pathways. J. Biol. Chem., 2010-11-12;286(4):2728-38. 2010-11-12 [PMID: 21075846] (ICC, Western Blot, Human)
• Poirier S, Mayer G, Poupon V, McPherson PS, Desjardins R, Ly K, Asselin MC, Day R, Duclos FJ, Witmer M, Parker R, Prat A, Seidah NG Dissection of the endogenous cellular pathways of PCSK9-induced low density lipoprotein receptor degradation: evidence for an intracellular route. J. Biol. Chem., 2009-07-27;284(42):28856-64. 2009-07-27 [PMID: 19635789] (ICC, Human)

Bioinformatics

• Gene Symbol: LDLR
• Uniprot:
  • P01130()