Anti-M6PR (cation independent) antibody [MEM-238] (ab8093)

Overview

  • Product nameAnti-M6PR (cation independent) antibody [MEM-238]
    See all M6PR (cation independent) primary antibodies
  • Description
    Mouse monoclonal [MEM-238] to M6PR (cation independent)
  • SpecificityCD222 antigen (human)
  • Tested applicationsSuitable for: Flow Cyt, IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Non Human Primates
  • Immunogen

    Vaccinia-M6PR/iGFIIR

  • EpitopeRecognizes an epitope between domains 2 and 5.
  • General notes


    CD222 is a 250kDa transmembrane protein with a short cytoplasmic tail containing an internalization signal. CD222 was originally identified as a receptor for IGFII and M6P-containing proteins (e.g. lysosomal hydrolases). Lysosomal enzymes are sorted to lysosomes via CD222 either from the Golgi, where the enzymes acquire M6P, or from the extracellular space. The majority of CD222 molecules (approximately 90-95%) are located intracellularly, only 5-10% is present on the cell membrane. The internalization rate seems to be enhanced by ligand induced dimerization of CD222 as well as by phosphorylation of its cytoplasmic serine. CD222 is also a receptor for TGFbeta latency associated peptide (LAP), proliferin and may bind several molecules independently of M6P, including plasminogen, CD87 or retinoic acid. It is involved in activation of latent TGFbeta [PROW].

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 15mM Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity>95% by SDS-PAGE
  • Purification notesPurified from ascites using protein A affinity chromatography.
  • Primary antibody notesCD222 is a 250kDa transmembrane protein with a short cytoplasmic tail containing an internalization signal. CD222 was originally identified as a receptor for IGFII and M6P-containing proteins (e.g. lysosomal hydrolases). Lysosomal enzymes are sorted to lysosomes via CD222 either from the Golgi, where the enzymes acquire M6P, or from the extracellular space. The majority of CD222 molecules (approximately 90-95%) are located intracellularly, only 5-10% is present on the cell membrane. The internalization rate seems to be enhanced by ligand induced dimerization of CD222 as well as by phosphorylation of its cytoplasmic serine. CD222 is also a receptor for TGFbeta latency associated peptide (LAP), proliferin and may bind several molecules independently of M6P, including plasminogen, CD87 or retinoic acid. It is involved in activation of latent TGFbeta [PROW].
  • ClonalityMonoclonal
  • Clone numberMEM-238
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8093 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. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IP Use a concentration of 10 µg/ml.
WB Use at an assay dependent concentration. Use under non reducing condition.
ICC/IF Use at an assay dependent concentration. Used at a concentration of 5 ug/ml for 1 hr on HEK cells (see Abreview for further information).

Target

  • FunctionTransport 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 similaritiesBelongs to the MRL1/IGF2R family.
    Contains 1 fibronectin type-II domain.
  • DomainContains 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 localizationLysosome 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

Anti-M6PR (cation independent) antibody [MEM-238] images

  • ab8093 at 5µg/ml staining human HEK cells by immunocytochemistry. The cells were fixed with paraformaldehyde and incubated with the antibody for 1 hour. Bound antibody was detected using a goat anti-mouse IgG Alexa-Fluor ® 568. In the confocal image ab8093 labelling in red shows a distribution consistent with the location of the trans-Golgi network and lysosomes. Blue nuclear counterstain is present.

    This image is courtesy of an Abreview submitted by Randal Moldrich on 31 March 2006.

    See Abreview

  • ab8093 staining Mannose 6 Phosphate Receptor (Cation independent) in human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde in PBS, permeabilized with 0.1% Triton X-100 and blocked with 2% BSA for 30 minutes at 25°C . Samples were incubated with primary antibody (1/300 in 2%BSA + 0.1% Triton X-100) for 2 hours at 25°C. An Alexa Fluor® 488-conjugated Goat polyclonal to mouse IgG (dilution 1/2000) was used as secondary antibody.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab8093 (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 (ab8093, 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-M6PR (cation independent) antibody [MEM-238] (ab8093)

This product has been referenced in:
  • Boonyaratanakornkit J  et al. Alix serves as an adaptor that allows human parainfluenza virus type 1 to interact with the host cell ESCRT system. PLoS One 8:e59462 (2013). ICC/IF ; Human . Read more (PubMed: 23527201) »
  • Schomacker H  et al. The C proteins of human parainfluenza virus type 1 block IFN signaling by binding and retaining Stat1 in perinuclear aggregates at the late endosome. PLoS One 7:e28382 (2012). ICC/IF . Read more (PubMed: 22355301) »

See all 8 Publications for this product

Product Wall

I have issued a free of charge replacement with the order number for ab32815.

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Should you still be experiencing difficulties, or i...

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Thank you, I received the data this time. For a protocol modification, I suggest trying preparing a sample by heating at 60C for 10 minutes instead of 95 for 5 minutes to minimize aggregation of the trans-membrane domains. It is strange that both antib...

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Thank you for bringing this to our attention. I will need a few more details of your protocol. I was not able to open the file you sent.

Can you please tell me how you prepared your lysates (did you use protease inhibitors), and how you heat...

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I'm glad that you have been seeing better results with blocking with BSA. If you are now having difficulty with non-specific bands, increasing the dilution of the primary and secondary antibody as well as adding 1 % BSA and 0.2% Tween 20 into the dilue...

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Thank you for contacting us. Having looked into the case a little I can see no reason why ab8093 should not be performing well with Western blotting or that ab2733 would be likely to perform any better. What might be helpful is if you were to share...

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I have attached an Excel file with information regarding all our current antibodies that have been tested in EM. I hope this is helpful. Please let me know if you have any further questions.

Thank you for your enquiry. I am working up an internal query to obtain a list of all EM-tested antibodies in our catalog. I will forward you this list as soon as possible. Thanks for your patience!

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (A549)
Specification A549
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step 5% Goat Serum, 0.5% BSA, 0.5% Gelatin as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jul 07 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - Triton X-100
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 11 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEK cells)
Specification HEK cells
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 5%
Username

Dr. Randal Moldrich

Verified customer

Submitted Mar 31 2006

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