Overview

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

    Synthetic peptide within Human LDL Receptor aa 650-750 (C terminal). The exact sequence is proprietary.
    Database link: P01130

  • Positive control
    • WB: HeLa and RAW264.7 cell lysate. HepG2 whole cell lysate. Mouse liver, lung and colon lysate. Human liver and plasma lysate. Flow Cyt: HeLa and U937 cells. ICC/IF: HeLa cells. IHC-P: Human hepatocellular carcinoma and liver tissue.
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

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 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

Images

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

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) 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 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) 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
    All lanes : 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

  • Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 0.4 µg/ml + HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

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

    Predicted band size: 95 kDa


    Exposure time: 3 minutes


    Blocking and diluting buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with the literature (PMID: 15199428, PMID: 8349823, PMID: 10906332, PMID: 24918045).

  • 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 (Human liver hepatocellular carcinoma cell line) cell lysate
    Lane 6 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : 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

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

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

    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
  • 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 minutes


    LDL Receptor contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • Overlay histogram showing 4% paraformaldehyde fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling LDL Receptor (red) with purified ab52818 at dilution of 1/70. The secondary antibody used was an 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.

  • Overlay histogram showing U937 (Human histiocytic lymphoma cell line) 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.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling 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 labeling LDL 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.

  • 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 counterstained with hematoxylin.

References

This product has been referenced in:
  • Zhu L  et al. Hepatocyte estrogen receptor alpha mediates estrogen action to promote reverse cholesterol transport during Western-type diet feeding. Mol Metab 8:106-116 (2018). WB . Read more (PubMed: 29331506) »
  • Ding W  et al. Osteopontin deficiency ameliorates Alport pathology by preventing tubular metabolic deficits. JCI Insight 3:N/A (2018). Read more (PubMed: 29563333) »
See all 48 Publications for this product

Customer reviews and Q&As

1-10 of 20 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Liver
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 23 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Liver
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 23 2018

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

Submitted Dec 06 2013

1-10 of 20 Abreviews or Q&A

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