Anti-LDL Receptor antibody [EP1553Y] (ab52818)

Overview

  • Product nameAnti-LDL Receptor antibody [EP1553Y]
    See all LDL Receptor primary antibodies
  • Description
    Rabbit monoclonal [EP1553Y] to LDL Receptor
  • Tested applicationsSuitable 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.

     

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to use products containing Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com

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

  • FunctionBinds 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 diseaseDefects 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 similaritiesBelongs 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 localizationCell 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 minutes
    LDL 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:
  • de Gonzalo-Calvo D  et al. Intratumor cholesteryl ester accumulation is associated with human breast cancer proliferation and aggressive potential: a molecular and clinicopathological study. BMC Cancer 15:460 (2015). WB ; Human . Read more (PubMed: 26055977) »
  • Marshall SM  et al. Reduction of VLDL secretion decreases cholesterol excretion in niemann-pick C1-like 1 hepatic transgenic mice. PLoS One 9:e84418 (2014). WB ; Mouse . Read more (PubMed: 24404162) »

See all 23 Publications for this product

Product Wall

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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Thanks for your email.
I believe this replacement order has been shipped and delivered on Tuesday July 18. It was on Abcam order reference number 1128916 which contained a vial of ab61392.
I hope this is helpful. Please let me know if you hav...

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab61392 and one vial of ab52818.

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