Overview

  • Product name
    Anti-LDL Receptor antibody
    See all LDL Receptor primary antibodies
  • Description
    Chicken polyclonal to LDL Receptor
  • Host species
    Chicken
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Recombinant fragment
    Predicted to work with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment, corresponding to amino acids 29-205 of Human LDL Receptor.

  • Positive control
    • LDLR transfected 293 cell lysate.

Associated products

Applications

Our Abpromise guarantee covers the use of ab14056 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
WB 1/1000. Detects a band of approximately 68, 100 kDa.
This antibody has been tested on recombinant protein only. We have not tested it on endogenous protein.

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

  • Transiently transfected 293 cells probed with ab14056. Transiently transfected 293 cells probed with ab14056.
  • Anti-LDL Receptor antibody (ab14056) at 1/5000 dilution + CHO-cells transfected with human LDLR at 20 µg

    Secondary
    Goat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 120,150 kDa
    why is the actual band size different from the predicted?


    Exposure time: 2 minutes

    See Abreview

References

This product has been referenced in:
  • Sato S  et al. Anti-fibrotic efficacy of nintedanib in pulmonary fibrosis via the inhibition of fibrocyte activity. Respir Res 18:172 (2017). Read more (PubMed: 28915889) »
  • Tong JG  et al. Spatial and temporal epithelial ovarian cancer cell heterogeneity impacts Maraba virus oncolytic potential. BMC Cancer 17:594 (2017). Read more (PubMed: 28854921) »
See all 21 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Western blot
Sample
Human Recombinant protein (CHO-cells transfected with human LDLR)
Loading amount
20 µg
Specification
CHO-cells transfected with human LDLR
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Feb 22 2007

Answer

Thank you for your enquiry. For ab14056, this was tested against a fusion protein corresponding to amino acids 29 to 205 of the human LDL receptor. Thus, the bands seen will be much smaller than for the full-length protein. Native LDLR will run as stated on the ab30532 datasheet, between 100 kDa and 160 kDa. It does look like you are getting approximately the same bands on your blots, only that the higher band is much more heavily stained by ab30532 than ab14056. The band marked B in each blot should be the unglycosylated protein, the band marked A I assume is fully glycosylated, and it appears the lower band is non-specific. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your email. Neither of these antibodies have been tested yet for application in FACS. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase one of these products, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 Abcam Points will be offered for an image). Please contact us again if you have any additional questions.

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Answer

Thank you for your enquiry. I have been in touch with the source of this antibody and I have determined that neutralisation is not recommended as an application for this antibody. Although mouse and human LDL-r have good homology in sequence and the antibody is polyclonal, so far it has not been shown to bind to the ligand binding site of the receptor with inhibitory properties. Furthermore it may not lock the site completely. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your enquiry and for taking your time to fill in the Questionnaire. We are very sorry to hear that you are having problem with this antibody. Could you please let us know the batch number you have received? It may well be that the antibody has just gone off during shipping and/or storage. We would also be grateful if you can specify for how long you incubated the samples with the primary antibody. We normally suggest 1-2 hrs incubation at room temperature or overnight at 4oC. Have you cultured the cells on the plastic coverslips? Could we ask if you have coated the coverslips? This is very important to make sure that the cultured cells keep their morphological, biochemical and molecular characteristics. Alternatively, you can stain freshly prepared cytospin preparations. We look forward to hearing from you soon.

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Question

1. Please describe the problem (high background, wrong band size, more bands, no band etc)no signal not even a weak!! 2. On what material are you testing the antibody in WB? • Species? mouse • Cell extract/ Nuclear extract? cell lysate • Purified protein? • Recombinant protein? endogeneous 3. How much protein did you load? 50 µg • How did you prepare the lysate for the analysis (protease inhibitors etc)? (Triton 0,5% + protease inhibitors) • Did you heat the samples? 95°C/5 min 1 x reducing conditions/ 2 x non reducing conditions 4. Primary Antibody • Specification (in which species was it raised against)? Chicken anti LDL-Receptor (ab14056) • At what dilution(s) have you tested this antibody? 1/5000 (recommended in your subscribtion, 1:2500, 1:2000 • Incubation time, wash step? 4°C/over night 5. Secondary Antibody • Specification (in which species was it raised against)? Anti chicken peroxidase conjugated (ab6877) • At what dilution(s) have you tested this antibody? 1/2500, 1/5000, 1/10000 • Incubation time, wash step? 1-2h at room temperature • Do you know whether the problems you are experiencing come from the secondary? 6. What detection method are you using? ECL, chemiluminescence 7. Optimization attempts • How many times have you tried the Western? Three times and 36 samples!!! • Do you obtain the same results every time e.g. are background bands always in the same place? • What steps have you altered? 8. Did you apply positive and negative controls along with the samples? Please specify. The system works in generally, I detected on the same membranes a 150 kDa Protein; with your antibody there is no signal, no background and no unspecific band; the secondary antibody was changed into an alkaline phosphatase conjugated anti chicken and there were no signals. I

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Answer

I'm sorry to hear ab14056 is giving your customer problems. I have had a good look at the protocol you sent and it looks fine, I would recommend sending him free of charge a new vial to try out. I hope this helps, please let me know if you need further advice,

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Answer

Thank you for your email, and I am very pleased to hear that ab14056 is working out for you in both Western blotting and IHC. I suggest that you submit a review (and get a gift certificate to Amazon.com in return!) - just go to the online datasheet for ab14056, click on the reviews tab and then click on add a review. I do apologize for the 2 ul shortage that you received; we do strive to be absolutely 100% accurate when aliquoting our antibodies. If you have any further questions, please don't hesitate to contact me again.

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Answer

Thank you very much for your patience. I have just received the following information regarding this antibody: ab14056 was raised against a protein domain of 176 amino acids at the N-terminus and is expected to work in IHC. However, this has not yet been tested. To our knowledge ab14056 has only been tested for application in Western blotting and has not been tested in other applications. There was a mistake on our online datasheet which I have since corrected and I do apologize for this. If you have purchased ab14056 and have found it to not work in IHC, I will gladly refund you. Please let me know if you have any further questions.

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