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RecombinantRabMAb

Recombinant Anti-ORP1 antibody [EPR8646] (ab131165)

Submit a review Submit a question References (10)
Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
  • Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
  • Flow Cytometry (Intracellular) - Anti-ORP1 antibody [EPR8646] (ab131165)
  • Immunocytochemistry/ Immunofluorescence - Anti-ORP1 antibody [EPR8646] (ab131165)
  • Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
  • Immunocytochemistry/ Immunofluorescence - Anti-ORP1 antibody [EPR8646] (ab131165)
  • Flow Cytometry (Intracellular) - Anti-ORP1 antibody [EPR8646] (ab131165)
  • Anti-ORP1 antibody [EPR8646] (ab131165)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR8646] to ORP1
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-ORP1 antibody [EPR8646]
    See all ORP1 primary antibodies

  • Description

    Rabbit monoclonal [EPR8646] to ORP1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WBmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, 293T, U87-MG cell lysates; Mouse brain and heart lysates; Rat brain lysates Flow Cyt (intra): A549 cells ICC/Flow Cyt (intra): A549 and U87-MG cells

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR8646

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab131165 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 (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/200.
WB
1/1000 - 1/10000. Detects a band of approximately 50-108 kDa (predicted molecular weight: 108 kDa).
Notes
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/200.
WB
1/1000 - 1/10000. Detects a band of approximately 50-108 kDa (predicted molecular weight: 108 kDa).
Application notes
Is unsuitable for IP.

Target

Images

  • Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
    Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
    All lanes : Anti-ORP1 antibody [EPR8646] (ab131165) at 1/10000 dilution (Purified)

    Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
    Lane 2 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysates
    Lane 3 : Mouse brain lysates
    Lane 4 : Mouse heart lysates
    Lane 5 : Rat brain lysates
    Lane 6 : Rat heart lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 108 kDa
    Observed band size: 108,50 kDa why is the actual band size different from the predicted?



    Two mRNA variants produce a 108 kDa long ORP1 (ORP1L) and a 50 KDa short ORP1 (ORP1S) separately, which is consistent with what has been described in PMID: 12631712.

  • Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
    Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
    Anti-ORP1 antibody [EPR8646] (ab131165) at 1/1000 dilution + 293T (Human embryonic kidney epithelial cell) whole cell lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 108 kDa
    Observed band size: 108,50 kDa why is the actual band size different from the predicted?



    Two mRNA variants produce a 108 kDa long ORP1 (ORP1L) and a 50 KDa short ORP1 (ORP1S) separately, which is consistent with what has been described in PMID: 12631712.

  • Flow Cytometry (Intracellular) - Anti-ORP1 antibody [EPR8646] (ab131165)
    Flow Cytometry (Intracellular) - Anti-ORP1 antibody [EPR8646] (ab131165)

    Intracellular Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling ORP1 with purified ab131165 at 1/60 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

     

  • Immunocytochemistry/ Immunofluorescence - Anti-ORP1 antibody [EPR8646] (ab131165)
    Immunocytochemistry/ Immunofluorescence - Anti-ORP1 antibody [EPR8646] (ab131165)
    Immunocytochemistry/ Immunofluorescence analysis of A549 (Human lung carcinoma epithelial cell) cells labeling ORP1 with purified ab131165 at 1:200 dilution (2.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
    Western blot - Anti-ORP1 antibody [EPR8646] (ab131165)
    All lanes : Anti-ORP1 antibody [EPR8646] (ab131165) at 1/1000 dilution (unpurified)

    Lane 1 : A549 cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : U87-MG cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 108 kDa
    Observed band size: 50-108 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-ORP1 antibody [EPR8646] (ab131165)
    Immunocytochemistry/ Immunofluorescence - Anti-ORP1 antibody [EPR8646] (ab131165)

    ab131165 (unpurified) staining ORP1 in U87-MG (human glioblastoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000.

    DAPI was used as a nuclear counterstain and the negative control was PBS only.

  • Flow Cytometry (Intracellular) - Anti-ORP1 antibody [EPR8646] (ab131165)
    Flow Cytometry (Intracellular) - Anti-ORP1 antibody [EPR8646] (ab131165)

    ab131165 (unpurified) at 1/10 dilution staining ORP1 in permeabilized A549 cells by intracellular flow cytometry (red). Rabbit IgG negative control (green). 

     

     

  • Anti-ORP1 antibody [EPR8646] (ab131165)
    Anti-ORP1 antibody [EPR8646] (ab131165)

References (10)

Publishing research using ab131165? Please let us know so that we can cite the reference in this datasheet.

ab131165 has been referenced in 10 publications.

  • Boutry M & Kim PK ORP1L mediated PI(4)P signaling at ER-lysosome-mitochondrion three-way contact contributes to mitochondrial division. Nat Commun 12:5354 (2021). PubMed: 34504082
  • Singhal A  et al. 2-Hydroxypropyl-gamma-cyclodextrin overcomes NPC1 deficiency by enhancing lysosome-ER association and autophagy. Sci Rep 10:8663 (2020). PubMed: 32457374
  • Zhao K  et al. Oxysterol-binding protein-related protein 1 variants have opposing cholesterol transport activities from the endolysosomes. Mol Biol Cell 31:793-802 (2020). PubMed: 32023146
  • Dong J  et al. Allosteric enhancement of ORP1-mediated cholesterol transport by PI(4,5)P2/PI(3,4)P2. Nat Commun 10:829 (2019). PubMed: 30783101
  • Atakpa P  et al. IP3 Receptors Preferentially Associate with ER-Lysosome Contact Sites and Selectively Deliver Ca2+ to Lysosomes. Cell Rep 25:3180-3193.e7 (2018). PubMed: 30540949
  • Di Mattia T  et al. Identification of MOSPD2, a novel scaffold for endoplasmic reticulum membrane contact sites. EMBO Rep 19:N/A (2018). PubMed: 29858488
  • Zhao K & Ridgway ND Oxysterol-Binding Protein-Related Protein 1L Regulates Cholesterol Egress from the Endo-Lysosomal System. Cell Rep 19:1807-1818 (2017). PubMed: 28564600
  • Martínez-Redondo V  et al. Peroxisome Proliferator-activated Receptor ? Coactivator-1 a Isoforms Selectively Regulate Multiple Splicing Events on Target Genes. J Biol Chem 291:15169-84 (2016). WB ; Mouse . PubMed: 27231350
  • Cianciola NL  et al. Adenovirus RIDa uncovers a novel pathway requiring ORP1L for lipid droplet formation independent of NPC1. Mol Biol Cell 24:3309-25 (2013). WB ; Human . PubMed: 24025716
  • Alpy F  et al. STARD3 or STARD3NL and VAP form a novel molecular tether between late endosomes and the ER. J Cell Sci 126:5500-12 (2013). PubMed: 24105263

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