使用敲除细胞株进行验证RabMAb

Anti-LRP1抗体[EPR3724] (ab92544)

概述

  • 产品名称Anti-LRP1抗体[EPR3724]
    参阅全部 LRP1 一抗
  • 描述
    兔单克隆抗体[EPR3724] to LRP1
  • 经测试应用适用于: IHC-P, WB, IP, Flow Cyt, ICC/IF, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Pig
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human LRP1 aa 4450 to the C-terminus.

  • 阳性对照
    • WB: PMBC and A549 cell lysates; mouse brain, heart, kidney and spleen tissue lysates; rat brain, heart, kidney or spleen tissue lysates; human fetal brain tissue lysates; pig liver and heart tissue lysates. IHC-P: Human liver, clear cell carcinoma, brain, lung and placenta tissues. ICC/IF: U87-MG cells. Flow Cyt: Jurkat cells. IP: A549 cells.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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.

     

    Alternative versions available:

    Anti-LRP1 antibody (Alexa Fluor® 488) [EPR3724] (ab195567)
    Anti-LRP1 antibody (Alexa Fluor® 647) [EPR3724] (ab195568)

    Anti-LRP1 antibody (HRP) [EPR3724] (ab195569)

    Anti-LRP1 antibody (Alexa Fluor® 555) [EPR3724] (ab206904)

应用

Our Abpromise guarantee covers the use of ab92544 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/5000 - 1/10000.

WB 1/50000. Predicted molecular weight: 85 kDa.

For unpurified use at 1/20000 - 1/50000.

IP 1/30.

For unpurified use at 1/50.

Flow Cyt 1/100.

For unpurified use at 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100.

For unpurified use at 1/5000 - 1/10000.

IHC-Fr Use at an assay dependent concentration.

靶标

  • 功能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.
  • 组织特异性Most abundant in liver, brain and lung.
  • 序列相似性Belongs to the LDLR family.
    Contains 22 EGF-like domains.
    Contains 31 LDL-receptor class A domains.
    Contains 34 LDL-receptor class B repeats.
  • 翻译后修饰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.
  • 细胞定位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
  • 数据库链接
  • 别名
    • 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

Anti-LRP1 antibody [EPR3724] 图像



  • Predicted band size : 85 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: LRP1 knockout HAP1 cell lysate (20 µg)
    Lane 3: A549 cell lysate (20 µg)
    Lane 4: Mouse heart tissue lysate (20 µg)


    Lanes 1 - 4: Merged signal (red and green). Green - ab92544 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab92544 was shown to specifically react with LRP1 when LRP1 knockout samples were used. Wild-type and LRP1 knockout samples were subjected to SDS-PAGE. ab92544 and ab8245 (loading control to GAPDH) were diluted at 1/5000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human clear cell carcinoma of the kidney tissue labelling LRP1 with purified ab92544 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of U87-MG cells labelling LRP1 with purified ab92544 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

  • All lanes : Anti-LRP1 antibody [EPR3724] (ab92544) at 1/50000 dilution (purified)

    Lane 1 : A549 cell lysate
    Lane 2 : Human fetal brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Flow Cytometry analysis of Jurkat cells labelling LRP1 with purified ab92544 at 1/100 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • ab92544 (purified) at 1/30 immunoprecipitating LRP1 in A549 cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-LRP1 antibody [EPR3724] (ab92544) at 1/50000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human brain tissue labelling LRP1 with unpurified ab92544.

  • ICC/IF image of LRP1 staining on rat mixed glia culture using unpurified ab92544 (1:200). The cells were fixed using paraformaldehyde. The cells were then permeabilised using 0.1% TritonX in 0.1% PBS. Non-specific protein was blocked using 10% donkey serum at 24°C for 1 hour. ab92544 was diluted (1/200) using 0.1% TritonX with 0.1% PBS and 10% donkey serum and the cells were incubated for 4 hours at 24°C. The secondary antibody used was donkey polyclonal to Rabbit IgG conjugated to Alexa Fluor® 488. DAPI was used to stain the nucleus.

    See Abreview

  • Overlay histogram showing Jurkat cells stained with unpurified ab92544 (red line). The cells were fixed with 80% methanol (5 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 (ab92544, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • All lanes : Anti-LRP1 antibody [EPR3724] (ab92544) at 1/50000 dilution (purified)

    Lane 1 : Pig liver tissue lysate
    Lane 2 : Pig heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human lung tissue labelling LRP1 with unpurified ab92544.

  • All lanes : Anti-LRP1 antibody [EPR3724] (ab92544) at 1/20000 dilution (unpurified)

    Lane 1 : Human PMBC lysate
    Lane 2 : A549 lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : Mouse kidney lysate
    Lane 6 : Mouse spleen lysate
    Lane 7 : Rat brain lysate
    Lane 8 : Rat heart lysate
    Lane 9 : Rat kidney lysate
    Lane 10 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size : 85 kDa
    Observed band size : 85 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human placenta tissue labelling LRP1 with unpurified ab92544.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling LRP1 with unpurified ab92544 at 1/100.

Anti-LRP1 antibody [EPR3724] (ab92544)参考文献

This product has been referenced in:
  • Bian Z  et al. Cd47-Sirpa interaction and IL-10 constrain inflammation-induced macrophage phagocytosis of healthy self-cells. Proc Natl Acad Sci U S A 113:E5434-43 (2016). Read more (PubMed: 27578867) »
  • Alata W  et al. Human apolipoprotein E ?4 expression impairs cerebral vascularization and blood-brain barrier function in mice. J Cereb Blood Flow Metab 35:86-94 (2015). Read more (PubMed: 25335802) »

See all 18 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (sarcoma tumor)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification sarcoma tumor
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 21°C
Fixative 10% NBF
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Abcam user community

Verified customer

提交于 Oct 25 2016

Application Western blot
Sample Guinea pig Tissue lysate - other (Brain)
Gel Running Conditions Reduced Denaturing (7.5)
Loading amount 1.25 µg
Specification Brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 May 04 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (liver)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification liver
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 18 2015

The LRP1 antibody ab92544 is raised against a peptide at the C terminus of isoform 1, the isoform described in the UniProt record for human LRP1 at this URL, http://www.uniprot.org/uniprot/Q07954. It is within the ICD domain and also the 85kDa subunit,...

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Application Immunocytochemistry/ Immunofluorescence
Blocking step PBS, 1% BSA, 10% goat serum, 0.1% TX-100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample Mouse Cell (primary embryonic epicardial)
Specification primary embryonic epicardial
Permeabilization Yes - 0.5% Triton X-100
Fixative Formaldehyde
Username

Abcam user community

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提交于 Jan 31 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: 23°C
Sample Mouse Tissue sections (brain)
Specification brain
Permeabilization Yes - 1%NaOH, 1%H2O2, 0.03% SDS solution
Fixative formalin->sucrose
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提交于 Jul 05 2013

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

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 Dec 14 2012

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...

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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 ...

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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 ...

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