Anti-M6PR (cation independent) antibody (ab32815)

Overview

  • Product nameAnti-M6PR (cation independent) antibodySee all M6PR (cation independent) primary antibodies ...
  • Description
    Rabbit polyclonal to M6PR (cation independent)
  • Tested applicationsWB, ICC, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Pig, Non Human Primates
  • Immunogen

    Full length native protein purified from adult Bovine liver tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab32815 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
WB 1/2000 - 1/3000. Detects a band of approximately 300 kDa (predicted molecular weight: 275 kDa).
ICC 1/50 - 1/500.
IP 1/1000 - 1/2000.
ICC/IF 1/50 - 1/500.
IHC-P 1/1000.

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. 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 images

  • Immunofluorescent analysis of Mannose 6 Phosphate Receptor (Cation independent) (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Mannose 6 Phosphate Receptor (Cation independent) antibody (ab32815) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunofluorescent analysis of Mannose 6 Phosphate Receptor (Cation independent)(green) showing staining in the cytoplasm and nucleus of MCF-7 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Mannose 6 Phosphate Receptor (Cation independent) antibody (ab32815) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunofluorescent analysis of Mannose 6 Phosphate Receptor (Cation independent)  (green) showing staining in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Mannose 6 Phosphate Receptor (Cation independent) antibody (ab32815) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Ab32815 staining human normal left ventricle of heart. Staining is localized to lysosome and lysosomal membrane.
    Left panel: with primary antibody duluted at 1:1000. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplifi
  • ICC/IF image of ab32815 stained HeLa cells. The cells were 4% formaldehyde 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 (ab32815, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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 for Anti-M6PR (cation independent) antibody (ab32815)

This product has been referenced in:
  • Ooms K  et al. Development of a recombinant antibody to target peptides and proteins to sialoadhesin-expressing macrophages. BMC Biotechnol 13:33 (2013). ICC/IF ; Pig . Read more (PubMed: 23575465) »
  • Laufman O  et al. The COG complex interacts directly with Syntaxin 6 and positively regulates endosome-to-TGN retrograde transport. J Cell Biol 194:459-72 (2011). Read more (PubMed: 21807881) »

See all 11 Publications for this product

Product Wall

Thank you for your reply.

If you would prefer, I would be happy to issue ab32815 as a free of charge replacement for ab129923.

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. The one protocol modification that might have an effect on the background stain would be to substitute eithe...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Pig Cell (primary alveolar macrophage)
Specification primary alveolar macrophage
Fixative Paraformaldehyde
Permeabilization Yes - TX-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
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Verified customer

Submitted Feb 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (Membrane fraction of Hela Cells)
Loading amount 100000 cells
Specification Membrane fraction of Hela Cells
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Abcam user community

Verified customer

Submitted Sep 12 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela Cells)
Specification Hela Cells
Fixative Methanol
Permeabilization No
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Verified customer

Submitted Aug 11 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (lacrimal gland)
Loading amount 25 µg
Specification lacrimal gland
Gel Running Conditions Reduced Denaturing
Blocking step Rockland Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C
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Abcam user community

Verified customer

Submitted Jul 10 2008

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab32815 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the ...

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