Overview

  • Product name
    Anti-LDL Receptor antibody
    See all LDL Receptor primary antibodies
  • Description
    Rabbit polyclonal to LDL Receptor
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Synthetic peptide:

    SVADTKGVKRRTL

    , corresponding to amino acids 499-511 of Mouse LDL Receptor

  • Positive control
    • Rat placenta, mouse liver, RAW 264.7 cell lysate

Applications

Our Abpromise guarantee covers the use of ab30532 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
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-Fr Use at an assay dependent concentration. Used at a dilution of 1/200 for 1 hr on mouse aortic root (see Abreview for further details).
IHC-P 1/400.
WB 1/200. Predicted molecular weight: 95 kDa.

The protein is highly glycosylated through N- and O-linkages and thus migrates at 100 kDa to 160 kDa on SDS-PAGE.

ICC/IF 1/100 - 1/400.

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

  • ab30532 staining LDL receptor in human lung parenchyma.
    Left panel: with primary antibody at 1/400. Right panel: isotype control.
    Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • ICC/IF image of ab30532 stained HepG2 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 (ab30532, 1/400) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

References

This product has been referenced in:
  • David M  et al. Identification and characterization of highly versatile peptide-vectors that bind non-competitively to the low-density lipoprotein receptor for in vivo targeting and delivery of small molecules and protein cargos. PLoS One 13:e0191052 (2018). WB, Flow Cyt . Read more (PubMed: 29485998) »
  • Liu MX  et al. Dicer1/miR-29/HMGCR axis contributes to hepatic free cholesterol accumulation in mouse non-alcoholic steatohepatitis. Acta Pharmacol Sin 38:660-671 (2017). Read more (PubMed: 28112179) »
See all 31 Publications for this product

Customer reviews and Q&As

1-10 of 25 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Gel Running Conditions
Non-reduced Denaturing (7.5)
Loading amount
25 µg
Specification
liver
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 23 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (SCC61 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
48 µg
Specification
SCC61 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 21 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293T, HepG2, MEF)
Loading amount
20 µg
Specification
293T, HepG2, MEF
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7% · Temperature: 20°C

Mr. Dongil Kim

Verified customer

Submitted Jan 16 2013

Answer


As we discussed, I will send you a positive control free of charge: ab7187 (RAW 264.7 whole cell lysate). This cell lysate gave a dark lower molecular weight band, the identity of which is unknown, along with bands at 110, 130, and 160 kDa corresponding with LDL receptor in different glycosylation states. In order to obtain a signal, we had to load a lot protein for rat placenta, mouse liver, and for RAW 264.7 cells (60 ug). Thus, the LDL receptor may not be present in very high amounts in your sample as well, so you may want to increase the amount that you are loading.

If the antibody does not detect bands in the 100-160 kDa range for the RAW 264.7 cell lysate, then the antibody is most likely not properly recognizing its target as we guarantee it will. However, if these bands are detected, then we know that the issue is not with the antibody, but is more likely that there is a very low amount of LDL receptor in your samples. If this is the case, I recommend loading more protein from your samples and using a higher primary antibody concentration.

Further western blot troubleshooting tips are given through the link below:
https://www.abcam.com/index.html?pageconfig=resource&rid=11352

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

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 with the order number xxx for another lot af ab30532.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

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 have any questions.

Read More

Answer

Thank you for contacting Abcam.

I have attached a western blot image showing that depending on the sample type that you are probing, you will get a number of different bands. Therefore, when running your western blot, you should expect to see number of bands at varying at varying molecular weights.

If there is anything else I can help you with, please let me know.

Read More

Question
Answer

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.

To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Question

Yes, please send us another aliquot of the 3-NT and anti LDL receptor antibody.
Perhaps we were just not lucky with that particular aliquot.
thanks, Jacob Raber
________________________________________
From: us.technical@abcam.com [us.technical@abcam.com]
Sent: Monday, July 16, 2012 6:57 AM
To: Jacob Raber
Subject: Reply from Abcam to your enquiry regarding ab30532, ab52818, ab61392 [CCE3956992]
Dear Dr Raber,
We have an answer to your inquiry:
Thanks for your reply.
Again I apologize for the trouble finding antibodies to 3-nitrotyrosine & the human LDL receptor. The antibodies you have ordered so far have very few
complaints and are ordered relatively frequently so it is certainly an unusual situation. Perhaps you would be interested in trying ab110282, another
antibody to 3-nitrotyrosine? If you are let me know and I can send it out as a replcement for PO 1632487.
Regarding ab52818 to LDLR, this antibody is also ordered relatively frequently and yours is the only complaint in the last 2 years. Perhaps it would
be worth just trying another vial of this same antibody? Although I can certainly send any antibody you would like to try to replace ab52818 when
I receive the PO information.
Help us improve our service. Rate your experience with us today. https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3956992
Your original inquiry to Abcam:
-------------------------------------------
That is correct; however, the replacement antibody did not work either. As you requested a PO and that antibody was not ordered,
I did not include it in the last e-mail. What we need is a antibody against 3-nitrotyrosine (3-NT) and one against the human LDL
receptor that both work for western blot analysis. We have no preference as long as they work for western blots. I cc Nicole in
our dept office to please share the PO for ab61392.
thanks again, Jacob Raber
________________________________________
From: us.technical@abcam.com [us.technical@abcam.com]
Sent: Friday, July 13, 2012 5:37 PM
To: Jacob Raber
Subject: Reply from Abcam to your enquiry re: RE: Reply from Abcam to your enquiry re: RE: Reply from Abcam to your enquiry re:
RE: Post complaint survey [CCE3954962]
Dear Dr Raber,
We have an answer to your inquiry:
Thanks for sending along the purchase order information.
Based on our records, you received a replacement end of May for ab30532. This replacement antibody was ab52818. So I can send you
1 free antibody to replace vial of ab52818 that did not work as expected.
Also I'll need the purchase order information for ab61392 so I can process a replacement for this antibody.
As well, in your reply, please let me know which antibodies you would like me to send you as replacements.
Thanks again for the additional information.
Help us improve our service. Rate your experience with us today. https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3954962
Your original inquiry to Abcam:
-------------------------------------------
Please find enclosed the requested info, regards, Jacob Raber
________________________________________
From: us.technical@abcam.com [us.technical@abcam.com]
Sent: Thursday, July 12, 2012 8:40 AM
To: Jacob Raber
Subject: Reply from Abcam to your enquiry re: RE: Reply from Abcam to your enquiry re: RE: Post complaint survey [CCE3949019]
Dear Dr Raber,
We have an answer to your inquiry:
Thanks again for the follow up information.
Its unusual to have multiple antibodies not give the expected results as you appear to be experiencing so I am sorry for this trouble.
If you can provide the PO numbers or Abcam order reference numbers regarding these 3 antibodies, I would be happy to offer replacements of
any primary antibody, credits or refunds.
Help us improve our service. Rate your experience with us today. https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3949019
Your original inquiry to Abcam:
-------------------------------------------
Post complaint survey [CCE3944284]
For LDLR antibody, the problem has been lack of bands on the blots.
However, probes for B-actin and apoE have worked on the same blots that come up negative for LDLR, suggesting it is not a loading/transfer issue.
The tissues we are running include a negative control (LDLR knockout tissue) as well as wild-type tissue and tissue that over-expresses human LDLR.
As far as protein tyrosine nitration, previous blots on similar samples (brain tissue from same genotype/treatment group of mice) using a 3NT antibody
from Millipore worked in the past. (We decided to try the Abcam 3NT antibody b/c of the much lower price)
thanks
Lance
******************************
Lance A. Johnson, PhD
Postdoctoral Fellow
Department of Behavioral Neuroscience
Oregon Health & Science University
(503) 494-1431
******************************
________________________________________
From: Jacob Raber
Sent: Wednesday, July 11, 2012 8:55 AM
To: Lance Johnson
Subject: FW: Reply from Abcam to your enquiry re: RE: Post complaint survey [CCE3944284]
Lance,
please respond and cc me, thanks, Jacob
________________________________________
From: us.technical@abcam.com [us.technical@abcam.com]
Sent: Wednesday, July 11, 2012 8:54 AM
To: Jacob Raber
Subject: Reply from Abcam to your enquiry re: RE: Post complaint survey [CCE3944284]
Dear Dr Raber,
We have an answer to your inquiry:
Thanks for the additional experimental details.
Is the problem that there are no bands on the blots or high background/non-specific bands?
The samples are good positive control samples for the LDLR antibodies. Is there evidence that protein tyrosine nitration occurs in detectable levels in these samples as well?
Help us improve our service. Rate your experience with us today. https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3944284
Your original inquiry to Abcam:
-------------------------------------------
Details for Post complaint survey [CCE3936459]:
We used a range of similar Western blot conditions for the two anti-LDLR antibodies 30532 and 52818.
We tried dilutions of 1:200, 1:500, 1:1000, 1:2000, and 1:5000 for the primary antibodies and dilutions ranging from 1:500-1:10000
for a HRP-conjugated secondary antibody (santa cruz). Blots were washed in 1X PBS with 0.2% Tween and blocked in the same solution
plus 2% milk (also tried BSA for blocking).
As far as tissue preparation, we tried both a whole tissue (brain) homogenate (homogenized in RIPA buffer), as well as a specific
cell-membrane preparation since LDLR is a cell surface receptor (using 0.32 M sucrose and sequential centrifugation).
Thank you for your help
Lance
******************************
Lance A. Johnson, PhD
Postdoctoral Fellow
Department of Behavioral Neuroscience
Oregon Health & Science University
(503) 494-1431
******************************
________________________________________
From: Jacob Raber
Sent: Monday, July 09, 2012 6:43 PM
To: Lance Johnson
Cc: Lauren Yeiser
Subject: FW: Post complaint survey [CCE3936459]
Lance,
can you please respond and provide the requested info and cc me? cc Lauren as well as we probably will need to do this for the other
abs as well.
thanks, Jacob
________________________________________
From: us.technical@abcam.com [us.technical@abcam.com]
Sent: Monday, July 09, 2012 3:14 PM
To: Jacob Raber
Subject: RE: Post complaint survey [CCE3936459]
Dear Dr Raber,
We have an answer to your inquiry:
Thanks for your reply.
Sorry for any confusion here. My email to you was just regarding the survey you filled out regarding ab30532. At this time I am happy
to send a replacement for this antibody. Just let me know which catalog number you would like as a replacement.
Please provide more complete experimental and sample preparation details on the other antibodies you have recently purchased
(ab30532, ab52818 and ab61392) that are not working as expected and I will gladly review those as well. If I can not improve
your results then we can discuss replacements for those too.
Thanks in advance for the additional information.
Help us improve our service. Rate your experience with us today. https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3936459
Your original inquiry to Abcam:
-------------------------------------------
That would be great; the following 3 antibodies we just purchased did not work in our western blot analysis:
anti-LDLR antibodies 30532 and 52818
Anti-3-Nitrotyrosine antibody ab61392
thank you very much for the replacement antibodies. Please let me know if you need the shiipping info again.
best regards, Jacob
-------------------------------------------
Best regards,
Jeremy
Jeremy Kasanov, PhD
Scientific Support Specialist
Abcam Inc.
www.abcam.com
Abcam Customer Services and Technical Support Team
www.abcam.com/technical
[CCE3944284]
-------------------------------------------
Best regards,
Jeremy
Jeremy Kasanov, PhD
Scientific Support Specialist
Abcam Inc.
www.abcam.com
Abcam Customer Services and Technical Support Team
www.abcam.com/technical
[CCE3949019]
-------------------------------------------
Best regards,
Jeremy
Jeremy Kasanov, PhD
Scientific Support Specialist
Abcam Inc.
www.abcam.com
Abcam Customer Services and Technical Support Team
www.abcam.com/technical
[CCE3954962]
-------------------------------------------
Best regards,
Jeremy
Jeremy Kasanov, PhD
Scientific Support Specialist
Abcam Inc.
www.abcam.com
Abcam Customer Services and Technical Support Team
www.abcam.com/technical
[CCE3956992]

Read More
Answer

Thank you for sending along the additional PO information. I have located that order for ab61392 on our system.
I am happy to send out the replacement antibodies. Since a couple options have been discussed for these replacement antibodies, I just want to confirm that, for the replacements, you would like one vial each of ab61392 and ab52818.
Thanks!

Read More

Answer

Thanks for your reply.
Again I apologize for the trouble finding antibodies to 3-nitrotyrosine & the human LDL receptor. The antibodies you have ordered so far have very few complaints and are ordered relatively frequently so it is certainly an unusual situation. Perhaps you would be interested in trying ab110282, another antibody to 3-nitrotyrosine? If you are let me know and I can send it out as a replcement for PO 1632487.
Regarding ab52818 to LDLR, this antibody is also ordered relatively frequently and yours is the only complaint in the last 2 years. Perhaps it would be worth just trying another vial of this same antibody? Although I can certainly send any antibody you would like to try to replace ab52818 when I receive the PO information.

Read More

1-10 of 25 Abreviews or Q&A

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