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Immunofluorescence analysis of Mannose 6 Phosphate Receptor / CD222 was done on 70% confluent log phase HepG2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Mannose 6 Phosphate Receptor / CD222 (2G11) Mouse Monoclonal Antibody (MA1066) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Human, Non-human primate, Rat|
|Published species reactivity||Rabbit, Rat, Non-human primate, Hamster, Bovine, Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified bovine ~300 kDa CI-MPR.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1µg per 10^6 cells|
|Immunocytochemistry (ICC)||2-5 µg/ml|
|Immunofluorescence (IF)||2-5 µg/ml|
|Immunohistochemistry (PFA fixed) (IHC (PFA))||Assay Dependent|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-066 detects mannose 6-phospate receptor (MPR) from human, rat, monkey and bovine tissues. This antibody does not detect Chinese hamster ovary cell MPR.
MA1-066 has been successfully used in Western blot, immunofluorescence, EM immunocytochemistry and immunoprecipitation procedures. By Western blot, this antibody detects a ~300 kDa protein representing MPR in HeLa cell extract under non-reducing conditions. Immunofluorescence staining of MPR in HeLa cells with MA1-066 results in perinuclear staining.
MA1-066 immunogen is purified bovine ~300 kDa Cl-MPR. This antibody is shown to recognize an epitope in the extracellular domain of MPR.
Lysosomal enzymes containing one or two mannose 6-phosphate (man6P) moieties are moved about in the cell by two distinct but interconnected cycles by means of ~300 kDa cation independent mannose 6-phospate receptors (CI-MPR). The biosynthetic cycle refers to the MPR transport of newly synthesized lysosomal enzymes from the trans Golgi network to late endosomes or early lysosomes. The endocytic cycle transports extracellular lysosomal enzymes from the plasma membrane via clathrin-coated vesicles to early endosomes. The entire pool of MPRs cycles between these cellular compartments every 3 hours. The steady state distribution of MPR's is predominantly within late endosomes, fewer in the trans Golgi network and ~10 % at cell surface.
In addition to its man6P binding activity, the MPR contains a separate binding site for the type II insulin-like growth factor and is capable of binding both man6P and IGF-II simultaneously. An ~240 kDa soluble, truncated form, representing the extracellular domain of the protein, has also been found circulating in serum and is capable of binding both ligands.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Human Copper Chaperone Atox1 Translocates to the Nucleus but does not Bind DNA In Vitro.
MA1-066 was used in immunocytochemistry to study the nuclear translocation of Atox1.
|Kahra D,Mondol T,Niemiec MS,Wittung-Stafshede P||Protein and peptide letters (22:532)||2015|
EGFR-dependent phosphorylation of leucine-rich repeat kinase LRRK1 is important for proper endosomal trafficking of EGFR.
MA1-066 was used in immunocytochemistry to study the role of LRRK1 phosphorylation by EGFR in the correct endosomal trafficking of EGFR
|Ishikawa K,Nara A,Matsumoto K,Hanafusa H||Molecular biology of the cell (23:1294)||2012|
Intracellular degradation of low-density lipoprotein probed with two-color fluorescence microscopy.
MA1-066 was used in immunocytochemistry to study the intracellular degradation of low-density lipoprotein using two-color fluorescence microscopy
|Humphries WH,Fay NC,Payne CK||Integrative biology : quantitative biosciences from nano to macro (2:536)||2010|
A spatio-temporal analysis of matrix protein and nucleocapsid trafficking during vesicular stomatitis virus uncoating.
MA1-066 was used in immunocytochemistry to investigate the spatio-temporal trafficking of matrix protein and nucleocapsid during vesicular stomatitis virus uncoating
|Mire CE,White JM,Whitt MA||PLoS pathogens (6:null)||2010|
Myosin Vc is a molecular motor that functions in secretory granule trafficking.
MA1-066 was used in immunocytochemistry to study the molecular motor role of myosin Vc in secretory granule trafficking
|Jacobs DT,Weigert R,Grode KD,Donaldson JG,Cheney RE||Molecular biology of the cell (20:4471)||2009|
EHD3 regulates early-endosome-to-Golgi transport and preserves Golgi morphology.
MA1-066 was used in immunocytochemistry to study the role of EHD3 in the regulation of endosome-to-Golgi transport
|Naslavsky N,McKenzie J,Altan-Bonnet N,Sheff D,Caplan S||Journal of cell science (122:389)||2009|
Identification of compartments involved in mammalian subcellular trafficking pathways by indirect immunofluorescence.
MA1-066 was used in immunocytochemistry to evaluate methods for the compartment identification in subcellular trafficking
|Doody A,Putnam D||Methods in molecular medicine (127:127)||2006|
Lowe syndrome protein OCRL1 interacts with clathrin and regulates protein trafficking between endosomes and the trans-Golgi network.
MA1-066 was used in immunocytochemistry to study the cellular function of hosphatidylinositol 4,5-bisphosphate 5-phosphatase OCRL1.
|Choudhury R,Diao A,Zhang F,Eisenberg E,Saint-Pol A,Williams C,Konstantakopoulos A,Lucocq J,Johannes L,Rabouille C,Greene LE,Lowe M||Molecular biology of the cell (16:3467)||2005|
A novel GTPase-activating protein for ARF6 directly interacts with clathrin and regulates clathrin-dependent endocytosis.
MA1-066 was used in immunocytochemistry to study SMAP1, a novel GTPase-activating protein for Arf6 that directly binds clathrin and modulates clathrin-mediated endocytosis
|Tanabe K,Torii T,Natsume W,Braesch-Andersen S,Watanabe T,Satake M||Molecular biology of the cell (16:1617)||2005|
Intracellular processing and activation of membrane type 1 matrix metalloprotease depends on its partitioning into lipid domains.
MA1-066 was used in immunocytochemistry to investigate the mechanism of integral membrane type 1 matrix metalloprotease (MT1-MMP) activities.
|Mazzone M,Baldassarre M,Beznoussenko G,Giacchetti G,Cao J,Zucker S,Luini A,Buccione R||Journal of cell science (117:6275)||2004|
A role for GRIP domain proteins and/or their ligands in structure and function of the trans Golgi network.
MA1-066 was used in immunocytochemistry to study the functions of GRIP proteins or their ligands in the maintenance of trans Golgi network integrity.
|Yoshino A,Bieler BM,Harper DC,Cowan DA,Sutterwala S,Gay DM,Cole NB,McCaffery JM,Marks MS||Journal of cell science (116:4441)||2003|
Accumulation of tyrosinase in the endolysosomal compartment is induced by U18666A.
MA1-066 was used in immunocytochemistry to study the accumulation of tyrosinase in an endolysosomal compartment of melanocytes treated with 3-beta-(2-diethylamonoethoxy)-androstenone
|Hall AM,Krishnamoorthy L,Orlow SJ||Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society (16:149)||2003|
Overexpression of an ADP-ribosylation factor-guanine nucleotide exchange factor, BIG2, uncouples brefeldin A-induced adaptor protein-1 coat dissociation and membrane tubulation.
MA1-066 was used in immunocytochemistry to investigate the role of BIG2 during the membrane association of AP-1.
|Shinotsuka C,Yoshida Y,Kawamoto K,Takatsu H,Nakayama K||The Journal of biological chemistry (277:9468)||2002|
Human myosin-Vc is a novel class V myosin expressed in epithelial cells.
MA1-066 was used in immunocytochemistry to investigate the role of myosin-Vc as a novel class V myosin.
|Rodriguez OC,Cheney RE||Journal of cell science (115:991)||2002|
Functionally distinct pools of actin in secretory cells.
MA1-066 was used in immunocytochemistry to investigate the importance of cytoskeleton in the secretion of HCl in stomach
|Ammar DA,Nguyen PN,Forte JG||American journal of physiology. Cell physiology (281:C407)||2001|
Sorting of mannose 6-phosphate receptors mediated by the GGAs.
MA1-066 was used in immunocytochemistry to study the role of Golgi-localized and ARF-binding proteins in mannose 6-phosphate receptor sorting.
|Puertollano R,Aguilar RC,Gorshkova I,Crouch RJ,Bonifacino JS||Science (New York, N.Y.) (292:1712)||2001|
Expression of rab11a N124I in gastric parietal cells inhibits stimulatory recruitment of the H+-K+-ATPase.
MA1-066 was used in immunocytochemistry to investigate the role of RAB11a N124I for stimulatory recruitment of the hydrogen-potassium-ATPase in gastric parietal cells
|Duman JG,Tyagarajan K,Kolsi MS,Moore HP,Forte JG||The American journal of physiology (277:C361)||1999|
Receptor extracellular domains may contain trafficking information. Studies of the 300-kDa mannose 6-phosphate receptor.
MA1-066 was used in immunocytochemistry to study the role of receptor extracellular domains in protein trafficking
|Dintzis SM,Velculescu VE,Pfeffer SR||The Journal of biological chemistry (269:12159)||1994|
Modulatory role of phosphoinositide 3-kinase in gastric acid secretion.
MA1-066 was used in western blot to investigate the role of phosphoinositide 3-kinase in gastric acid secretion
|Mettler SE,Ghayouri S,Christensen GP,Forte JG||American journal of physiology. Gastrointestinal and liver physiology (293:G532)||2007|
Serum form of the rat insulin-like growth factor II/mannose 6-phosphate receptor is truncated in the carboxyl-terminal domain.
MA1-066 was used in western blot to investigate the proteolysis of mannose 6-phosphate receptor protein in rat serum
|MacDonald RG,Tepper MA,Clairmont KB,Perregaux SB,Czech MP||The Journal of biological chemistry (264:3256)||1989|
High-pressure freezing of isolated gastric glands provides new insight into the fine structure and subcellular localization of H+/K+-ATPase in gastric parietal cells.
MA1-066 was used in immunohistochemistry to show the subcellular localization of hydrogen/potassium-ATPase in gastric parietal cells
|Sawaguchi A,McDonald KL,Forte JG||The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (52:77)||2004|
Mechanism of constitutive export from the golgi: bulk flow via the formation, protrusion, and en bloc cleavage of large trans-golgi network tubular domains.
MA1-066 was used in immunohistochemistry to investigate constitutive Golgi complex export process .
|Polishchuk EV,Di Pentima A,Luini A,Polishchuk RS||Molecular biology of the cell (14:4470)||2003|
300 kDa mannose 6-phosphate receptor; cation-independent mannose-6-phosphate receptor; CD222; CI Man-6-P receptor; CI-MPR; CIMPR; IGF-II receptor; IGF2R; insulin-like growth factor 2 receptor; insulin-like growth factor II receptor; insulin-like growth factor receptor type 2; M6P-R; M6P/IGF2 receptor; M6P/IGF2R; M6PR; MPR 300; MPR1; MPRI
BOS_9999; CD222; CI-M6PR; CI-MPR; CIMPR; IGF2R; Igfr2; M6P; M6P-R; M6P/IGF2R; MPR 300; MPR1; MPR300; MPRI