Overview

  • Product name
    Anti-LRP1 antibody [5A6]
    See all LRP1 primary antibodies
  • Description
    Mouse monoclonal [5A6] to LRP1
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, IHC-Fr, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
  • Immunogen

    Recombinant fragment (Human), 85kDa cleaved fragment of Low Density LRP

  • Positive control
    • Ab28320 gave a positive signal in Western Blot in WI38 whole cell lysate. This antibody also gave a positive result in IHC in the following FFPE tissue: Human normal placenta.
  • General notes

    Western blot protocol advice:

    We do not recommend adding a reducing agent (e.g. DTT) to lysates or heating samples prior to loading on SDS-PAGE gels. We have used BSA as our blocking agent at 1%.

    This antibody clone is manufactured by Abcam. If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Applications

Our Abpromise guarantee covers the use of ab28320 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 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

WB Use a concentration of 1 µg/ml. Use under non reducing condition. Detects a band of approximately 85 kDa (predicted molecular weight: 85 kDa).

Abcam do not recommend adding a reducing agent (e.g. DTT) to lysates or heating samples prior to loading on SDS-PAGE gels.  We have used BSA as our blocking agent at 1%.

IHC-P Use a concentration of 10 µg/ml.
IHC-Fr Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    Endocytic receptor involved in endocytosis and in phagocytosis of apoptotic cells. Required for early embryonic development. Involved in cellular lipid homeostasis. Involved in the plasma clearance of chylomicron remnants and activated LRPAP1 (alpha 2-macroglobulin), as well as the local metabolism of complexes between plasminogen activators and their endogenous inhibitors. May modulate cellular events, such as APP metabolism, kinase-dependent intracellular signaling, neuronal calcium signaling as well as neurotransmission.
    Functions as a receptor for Pseudomonas aeruginosa exotoxin A.
  • Tissue specificity
    Most abundant in liver, brain and lung.
  • Sequence similarities
    Belongs to the LDLR family.
    Contains 22 EGF-like domains.
    Contains 31 LDL-receptor class A domains.
    Contains 34 LDL-receptor class B repeats.
  • Post-translational
    modifications
    Cleaved into a 85 kDa membrane-spanning subunit (LRP-85) and a 515 kDa large extracellular domain (LRP-515) that remains non-covalently associated. Gamma-secretase-dependent cleavage of LRP-85 releases the intracellular domain from the membrane.
    The N-terminus is blocked.
    Phosphorylated on serine and threonine residues.
    Phosphorylated on tyrosine residues upon stimulation with PDGF. Tyrosine phosphorylation promotes interaction with SHC1.
  • Cellular localization
    Cytoplasm. Nucleus. After cleavage, the intracellular domain (LRPICD) is detected both in the cytoplasm and in the nucleus and Cell membrane. Membrane, coated pit.
  • Information by UniProt
  • Database links
  • Alternative names
    • A2MR antibody
    • Alpha 2 macroglobulin receptor antibody
    • alpha 2MR antibody
    • Alpha-2-macroglobulin receptor antibody
    • APOER antibody
    • Apolipoprotein E receptor antibody
    • APR antibody
    • CD 91 antibody
    • CD91 antibody
    • CD91 antigen antibody
    • IGFBP3R antibody
    • LDL receptor related protein 1 antibody
    • Low density lipoprotein receptor related protein 1 antibody
    • Low density lipoprotein related protein 1 antibody
    • Low-density lipoprotein receptor-related protein 1 intracellular domain antibody
    • LRP 1 antibody
    • LRP 515 antibody
    • LRP 85 antibody
    • LRP antibody
    • LRP ICD antibody
    • LRP-1 antibody
    • LRP-515 antibody
    • LRP-85 antibody
    • Lrp1 antibody
    • LRP1 protein antibody
    • LRP1_HUMAN antibody
    • LRP1A antibody
    • LRP515 antibody
    • LRP85 antibody
    • LRPICD antibody
    • MGC88725 antibody
    • Prolow density lipoprotein receptor related protein 1 antibody
    • TbetaR V/LRP 1/IGFBP 3 receptor antibody
    • TbetaRV/LRP1/IGFBP3 receptor antibody
    • TGFBR 5 antibody
    • TGFBR5 antibody
    • Type V tgf beta receptor antibody
    see all

Images

  • IHC image of LRP1 staining in Human normal placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab28320, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Anti-LRP1 antibody [5A6] (ab28320) at 1 µg/ml + WI38 (Human lung fibroblast cell line) Whole Cell Lysate [Non-reduced] at 25 µg

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 85 kDa
    Observed band size: 85 kDa


    Exposure time: 20 minutes


    This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 1% Bovine Serum Albumin before being incubated with ab28320 overnight at 4°C.  Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.  Please note that no reducing agent was added to the cell lysate and the sample was not heated prior to loading onto the SDS-PAGE gel. 

  • ab28320 at 1/3000 + WI38 fibroblasts whole cell lysate

    Secondary
    HRP conjugated donkey anti-mouse polyclonal diluted 1/10000

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Predicted band size: 85 kDa
    Observed band size: 85 kDa


    Exposure time: 2 minutes


    50µg of human fibroblast whole cell lysate were loaded on to the gel and run under denaturing conditions.

    The membrane was blocked with 1% BSA for 1 hour at RT.

    The primary antibody was incubated for 12 hours at 4°C.

    See Abreview

  • ICC/IF image of LRP1 staining in rat mixed glia culture using ab28320 (1:500). The cells were fixed uisng paraformaldehyde and permeabilised using 0.3% TritonX in 0.1%PBS. The cells were then blocked using 10% donkey serum for 1 hour at 24°C. the Cells were then inubated with ab23020 for 4 hours at 24°C. The secondary antibody used was donkey polyclonal to anti mouse IgG conjuagted to Alexa Fluor 568.

    See Abreview

  • Overlay histogram showing HepG2 cells stained with ab28320 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28320, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HepG2 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Willoughby JLS  et al. Evaluation of GalNAc-siRNA Conjugate Activity in Pre-clinical Animal Models with Reduced Asialoglycoprotein Receptor Expression. Mol Ther 26:105-114 (2018). WB . Read more (PubMed: 28988716) »
  • Chen J  et al. SLAMF7 is critical for phagocytosis of haematopoietic tumour cells via Mac-1 integrin. Nature 544:493-497 (2017). WB . Read more (PubMed: 28424516) »
See all 19 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

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

Dr. coralie brifault

Verified customer

Submitted Sep 13 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Mixed glial culture)
Specification
Mixed glial culture
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Dec 11 2012

Answer

Thank you for getting back to me.

Your order number for your free of charge replacment antibody ab92544 is 1210558. You should be receiving the antibody tomorrow. Also, for your own records, it appears the PO number used for the original order (order 1186298) was XXXXX

Your review of both antibodies will be much appreciated, and you will also receive points for your reviews that you can use for future products or Amazon gift cards. A more in depth description of the Abreview program is below:
https://www.abcam.com/index.html?pageconfig=resource&rid=10332&viapagetrap=abreviews

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link:
https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer


I now have answer from the producing lab of ab28320. I can confirm that Immunogen is the 85kDa cleaved fragment of Low Density LRP and therefore it is not strictly c-terminal.
I hope the other antibody that binds a c-terminal epitope ab92544 is suitable for your customer’s experiments.
I wish a good weekend.

Read More

Answer

Please excuse the delay in my answer.
I am still waiting to hear back from the producing lab concerning the immunogen of ab28320.
But I am happy to confirm that ab92544 (Anti-LRP1 antibody [EPR3724]) binds to c-terminal (cytoplasmic) part of LRP1.
I hope this information is helpful and would like to thank you for your patience.

Read More

Answer

I hope you are well.
I am happy to confirm that the current lot in stock of ab28320 has a concentration of 7.6mg/ml.
I hope this information is helpful and wish you a good week.

Read More

Question
Answer

Thank you for contacting Abcam and I do apologize for the delay in this reply.

I don't have a specific protocol for flow cytometry. We do have FACS dilution suggested at 1:1500. I have also attached the general Abcam protocols booklet, which contains lots of useful information.

Once again, I do apologize for the delay and if there is anything else I can help you with, please let me know.

Read More

Question
Answer

Thank you for your phone call.

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab28320 in rat. This code will give you: 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for ratand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you very much for your interest in LRP1 antibodies for use in neutralizing assays.

As I had mentyioned on the phone, we have not yet tested any of our anti-LRP1 antibodies in this application. As such these would qualify for participation in our 100% Abreview testing discount program. I have included some information about this program below.

When a product has not been tested in an application, I can offer a discount off a future purchase if you buythe product, test it in the untested application or species and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free PRIMARY ANTIBODY OR VALUE OFF OF NEXT ORDER.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab28320 or another anti-LRP1 antibody in an neutralizing/blocking assay. We will then send a discount code. This code must be issued before purchasing so please wait for our reply before ordering.

2. Purchase either by phone, fax, or online (www.abcam.com).

3. Test in your application.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if it turns out to be unsuitable for your application, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
No
Blocking step
Serum as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Nov 08 2011

1-10 of 15 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up