Overview

  • Product name
    Anti-M6PR (cation independent) antibody [2G11]
    See all M6PR (cation independent) primary antibodies
  • Description
    Mouse monoclonal [2G11] to M6PR (cation independent)
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-Fr, IPmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, African green monkey
    Predicted to work with: Non human primatesDoes not react with: Hamster
  • Immunogen

    Purified bovine 300 kDa Mannose 6 Phosphate Receptor (Cation independent).

  • Epitope
    This antibody is shown to recognize an epitope in the extracellular domain of Mannose 6 Phosphate Receptor.
  • Positive control
    • In Flow Cytometry, this antibody gave a positive signal in A431 cells.
  • General notes

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab2733 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.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 5 - 10 µg/ml.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for WB.
  • Target

    • Function
      Transport of phosphorylated lysosomal enzymes from the Golgi complex and the cell surface to lysosomes. Lysosomal enzymes bearing phosphomannosyl residues bind specifically to mannose-6-phosphate receptors in the Golgi apparatus and the resulting receptor-ligand complex is transported to an acidic prelyosomal compartment where the low pH mediates the dissociation of the complex. This receptor also binds IGF2. Acts as a positive regulator of T-cell coactivation, by binding DPP4.
    • Sequence similarities
      Belongs to the MRL1/IGF2R family.
      Contains 1 fibronectin type-II domain.
    • Domain
      Contains 15 repeating units of approximately 147 AA harboring four disulfide bonds each. The most highly conserved region within the repeat consists of a stretch of 13 AA that contains cysteines at both ends.
    • Cellular localization
      Lysosome membrane. Colocalized with DPP4 in internalized cytoplasmic vesicles adjacent to the cell surface.
    • Information by UniProt
    • Database links
    • Alternative names
      • 300 kDa mannose 6 phosphate receptor antibody
      • 300 kDa mannose 6-phosphate receptor antibody
      • Cation independent mannose 6 phosphate receptor antibody
      • Cation-independent mannose-6-phosphate receptor antibody
      • CD222 antibody
      • CD222 antigen antibody
      • CI Man 6 P receptor antibody
      • CI Man-6-P receptor antibody
      • CI MPR antibody
      • CI-M6PR antibody
      • CI-MPR antibody
      • CIMPR antibody
      • IGF 2 receptor antibody
      • IGF 2R antibody
      • IGF II receptor antibody
      • IGF-II receptor antibody
      • IGF2 receptor antibody
      • Igf2r antibody
      • Insulin like growth factor 2 receptor antibody
      • Insulin like growth factor II receptor antibody
      • Insulin-like growth factor 2 receptor antibody
      • Insulin-like growth factor II receptor antibody
      • M6P R antibody
      • M6P/IGF2 receptor antibody
      • M6P/IGF2R antibody
      • M6PR antibody
      • mannose 6 phosphate receptor antibody
      • mannose 6 phosphate receptor, cation independent antibody
      • MPR 300 antibody
      • MPR300 antibody
      • MPRI antibody
      • MPRI_HUMAN antibody
      see all

    Images

    • ICC/IF image of ab2733 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab2733, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    • Overlay histogram showing HeLa cells stained with ab2733 (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 (ab2733, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
    • Immunofluorescent imaging of human cells (U2OS) with ab2733 confirms the specificity of this antibody, with the expected perinuclear vesicular staining of lysosomes. 

      IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes.  All blocking and incubation steps carried out at 37 degrees.

    • Immunostained frozen sections showing that mannose 6 phosphate receptor is localised in the outermost epithelial cell layer.

      This picture was kindly supplied as part of the review submitted by Marko Nykanen.

    • ab2733 positively staining formaldehyde fixed Human HEK 293 cells (red) in conjunction with goat anti mouse (Alexa 546). Nuclear staining was obtained using Hoechst.
      This image is an edited version of an image received courtesy of an Abreview submitted by Kun Liu on 19 September 2005. We do not have any further information relating to this image.

      See Abreview

    • ICC/IF image of ab2733 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 (ab2733, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 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:
    • Sapmaz A  et al. USP32 regulates late endosomal transport and recycling through deubiquitylation of Rab7. Nat Commun 10:1454 (2019). Read more (PubMed: 30926795) »
    • Koike S & Jahn R SNAREs define targeting specificity of trafficking vesicles by combinatorial interaction with tethering factors. Nat Commun 10:1608 (2019). Read more (PubMed: 30962439) »
    See all 92 Publications for this product

    Customer reviews and Q&As

    Filter by Application

    Filter by Species

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    1-10 of 15 Abreviews

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (MEFs)
    Permeabilization
    Yes - 0.1%Triton
    Specification
    MEFs
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 18 2019

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - TritonX100
    Specification
    HeLa
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 22°C
    Fixative
    Paraformaldehyde

    Dr. Jerome Cattin

    Verified customer

    Submitted Jun 19 2019

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    HeLa
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 11 2018

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Cow Cell (MDBK - Madin-Darby Bovine Kidney)
    Specification
    MDBK - Madin-Darby Bovine Kidney
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton X-100
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jun 13 2011

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK293)
    Specification
    HEK293
    Fixative
    100% EtOH
    Permeabilization
    Yes - 0.1% Triton X-100
    Blocking step
    Gelatin as blocking agent for 40 minute(s) · Concentration: 0.1% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jan 14 2011

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa cells)
    Specification
    HeLa cells
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% Triton X-100
    Blocking step
    Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 20°C

    Dr. Margaret Robinson

    Verified customer

    Submitted Dec 02 2010

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK293 cell)
    Specification
    HEK293 cell
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton X-100
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

    Abcam user community

    Verified customer

    Submitted Feb 04 2010

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (blastocyst)
    Specification
    blastocyst
    Fixative
    Formaldehyde
    Permeabilization
    Yes - tritonX-100
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted May 21 2009

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Rat2 Cells)
    Specification
    Rat2 Cells
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% Triton
    Blocking step
    BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Dec 12 2008

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Rat2 Cells)
    Specification
    Rat2 Cells
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% Saponin
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 37°C

    Abcam user community

    Verified customer

    Submitted Dec 01 2008

    1-10 of 15 Abreviews

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