Recombinant
RabMAb

Anti-LDL Receptor antibody [EP1553Y] (ab52818)

Overview

  • Product name
    Anti-LDL Receptor antibody [EP1553Y]
    See all LDL Receptor primary antibodies
  • Description
    Rabbit monoclonal [EP1553Y] to LDL Receptor
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human LDL Receptor (C terminal).

  • Positive control
    • HepG2 cells and cell lysate
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Applications

Our Abpromise guarantee covers the use of ab52818 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/250.
IHC-P 1/500.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

WB 1/5000. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa).
Flow Cyt 1/70 - 1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Binds LDL, the major cholesterol-carrying lipoprotein of plasma, and transports it into cells by endocytosis. In order to be internalized, the receptor-ligand complexes must first cluster into clathrin-coated pits. In case of HIV-1 infection, functions as a receptor for extracellular Tat in neurons, mediating its internalization in uninfected cells.
  • Involvement in disease
    Defects in LDLR are the cause of familial hypercholesterolemia (FH) [MIM:143890]; a common autosomal semi-dominant disease that affects about 1 in 500 individuals. The receptor defect impairs the catabolism of LDL, and the resultant elevation in plasma LDL-cholesterol promotes deposition of cholesterol in the skin (xanthelasma), tendons (xanthomas), and coronary arteries (atherosclerosis).
  • Sequence similarities
    Belongs to the LDLR family.
    Contains 3 EGF-like domains.
    Contains 7 LDL-receptor class A domains.
    Contains 6 LDL-receptor class B repeats.
  • Post-translational
    modifications
    N- and O-glycosylated.
    Ubiquitinated by MYLIP leading to degradation.
  • Cellular localization
    Cell membrane. Endomembrane system. Membrane > clathrin-coated pit. Found distributed from the plasma membrane to intracellular compartments.
  • Information by UniProt
  • Database links
  • Alternative names
    • FH antibody
    • FHC antibody
    • LDL R antibody
    • LDL receptor antibody
    • LDLCQ2 antibody
    • Ldlr antibody
    • LDLR_HUMAN antibody
    • Low Density Lipoprotein Receptor antibody
    • Low density lipoprotein receptor class A domain containing protein 3 antibody
    • Low density lipoprotein receptor familial hypercholesterolemia antibody
    • Low-density lipoprotein receptor antibody
    see all

Anti-LDL Receptor antibody [EP1553Y] images

  • All lanes : Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : HeLa treated with GW3965 for 8 hours at the final concentration of 5uM whole cell lysates
    Lane 3 : HeLa treated with GW3965 for 24 hours at the final concentration of 5uM whole cell lysates
    Lane 4 : Raw264.7 whole cell lysates
    Lane 5 : Raw264.7 treated with GW3965 for 8 hours at the final concentration of 5uM whole cell lysates
    Lane 6 : Raw264.7 treated with GW3965 for 24 hours at the final concentration of 5uM whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (HRP goat anti-rabbit IgG (H+L))

    Predicted band size : 95 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    Blocking buffer: 5% NFDM/TBST

    Diltuion buffer: 5% NFDM/TBST

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling LDL Receptor with purified ab52818 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor®594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei were counterstained with DAPI (blue).

    For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used. 

  • Immunohistochemical analysis of paraffin-embedded human liver sections labellingLDL Receptor with purified ab52818 at dilution of 1:500. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Overlay histogram showing 4% paraformaldehyde fixed HeLa cells labelling LDL Receptor (red) with purified ab52818 at dilution of 1/70. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • All lanes : Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/1000 dilution

    Lane 1 : mouse liver lysate
    Lane 2 : mouse lung lysate
    Lane 3 : mouse colon lysate
    Lane 4 : human liver lysate
    Lane 5 : HepG2 cell lysate
    Lane 6 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (HRP goat anti-rabbit IgG (H+L))

    Predicted band size : 95 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    Blocking buffer: 5% NFDM/TBST

    Diltuion buffer: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling LDL Receptor with ab52818 at 1/100 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.

  • Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/5000 dilution + Mouse liver at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 20000

    Predicted band size : 95 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM/TBST
  • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling LDL Receptor with ab52818 at 1/100 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). The sample was counter stained with hematoxylin.

  • ICC/IF image of ab52818 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52818, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/5000 dilution + HepG2 whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 95 kDa
    Observed band size : 140 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM/TBST
  • Overlay histogram showing U937 cells stained with ab52818 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52818, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/500 dilution + Human Plasma Total Protein Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 95 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 27 kDa,48 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutesLDL Receptor contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

References for Anti-LDL Receptor antibody [EP1553Y] (ab52818)

This product has been referenced in:
  • Pinkosky SL  et al. Liver-specific ATP-citrate lyase inhibition by bempedoic acid decreases LDL-C and attenuates atherosclerosis. Nat Commun 7:13457 (2016). Read more (PubMed: 27892461) »
  • Jensen KK  et al. Dose-dependent effects of siRNA-mediated inhibition of SCAP on PCSK9, LDLR, and plasma lipids in mouse and rhesus monkey. J Lipid Res 57:2150-2162 (2016). WB ; Mouse . Read more (PubMed: 27707816) »

See all 30 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (bEnd.3 cell line)
Permeabilization
Yes - 0.1% triton
Specification
bEnd.3 cell line
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde
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Submitted Oct 11 2017

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain Tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
Yes - 0.2% Triton X-100
Specification
Brain Tissue
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Submitted Feb 15 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.1% Tween 20
Specification
Brain
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Formaldehyde
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Submitted Nov 03 2016

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (8%)
Sample
Rat Cell lysate - whole cell (liver cells)
Specification
liver cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Dec 06 2013

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (8%)
Sample
Human Cell lysate - whole cell (liver cells)
Specification
liver cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 37°C
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Submitted Dec 06 2013

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
No
Sample
Human Cell (liver cells)
Specification
liver cells
Gating Strategy
CD24 POSITIVE
Preparation
Cell harvesting/tissue preparation method: single cell suspensionn
Sample buffer: 0.1%PBS-BSA
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Submitted Dec 06 2013

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
No
Sample
Rat Cell (liver cells)
Specification
liver cells
Preparation
Cell harvesting/tissue preparation method: single cell suspensionn
Sample buffer: 0.1%PBS-BSA
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Submitted Dec 06 2013

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
No
Sample
Mouse Cell (liver cells)
Specification
liver cells
Gating Strategy
CD24 POSITIVE
Preparation
Cell harvesting/tissue preparation method: single cell suspensionn
Sample buffer: 0.1%PBS-BSA
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Submitted Dec 06 2013

Application
Western blot
Sample
Rabbit Tissue lysate - whole (liver)
Loading amount
30 µg
Specification
liver
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: rt°C
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Submitted May 13 2013

Gracias por tu respuesta.

Acabo de crear un código de descuento para testar ab52818 mediante Western Blot con lisados de células de conejo.

El código es: ABTRIAL-EBDAM. Caduca el día 23-Mayo-2013.
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