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Anti-Moesin/MSN Antibody Picoband™ (monoclonal, 8D4)

Figure 2. IHC analysis of Moesin/MSN using anti-Moesin/MSN antibody (M00766-2).
Moesin/MSN was detected in a paraffin-embedded section of human gastric adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Moesin/MSN Antibody (M00766-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 3. IHC analysis of Moesin/MSN using anti-Moesin/MSN antibody (M00766-2).
Moesin/MSN was detected in a paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Moesin/MSN Antibody (M00766-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 4. IHC analysis of Moesin/MSN using anti-Moesin/MSN antibody (M00766-2).
Moesin/MSN was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Moesin/MSN Antibody (M00766-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 5. IHC analysis of Moesin/MSN using anti-Moesin/MSN antibody (M00766-2).
Moesin/MSN was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Moesin/MSN Antibody (M00766-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 6. IF analysis of Moesin/MSN using anti-Moesin/MSN antibody (M00766-2).
Moesin/MSN was detected in an immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-Moesin/MSN Antibody (M00766-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 7. Flow Cytometry analysis of U87 cells using anti-Moesin/MSN antibody (M00766-2).
Overlay histogram showing U87 cells stained with M00766-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Moesin/MSN Antibody (M00766-2, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 1. Western blot analysis of Moesin/MSN using anti-Moesin/MSN antibody (M00766-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HL-60 whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: monkey COS-7 whole cell lysates,
Lane 4: rat kidney tissue lysates,
Lane 5: mouse kidney tissue lysates,
Lane 6: human MCF-7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Moesin/MSN antigen affinity purified monoclonal antibody (Catalog # M00766-2) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Moesin/MSN at approximately 78 kDa. The expected band size for Moesin/MSN is at 78 kDa.

SPECIFICATION
CITATIONS
PROTOCOLS
BACKGROUND

Product Information
Application Applications Legend: WB=Western Blot IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin-embedded Sections) IHC-F=Immunohistochemistry (Frozen Sections) IF=Immunofluorescence FC=Flow Cytopmetry IC=Immunochemistry ICC=Immunocytochemistry E=ELISA IP=Immunoprecipitation DB=Dot Blot CHIP=Chromatin Immunoprecipitation FA=Fluorescence Assay IEM=Immuno electron microscopy EIA=Enzyme Immunoassay	WB, IHC, IF, ICC, FC
Primary Accession	P26038
Host	Mouse
Isotype	Mouse IgG1
Reactivity	Rat, Human, Mouse, Monkey
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-Moesin/MSN Antibody Picoband™ (monoclonal, 8D4) . Tested in FCM, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat, Monkey.
Reconstitution	Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Additional Information
Gene ID	4478
Other Names	Moesin, Membrane-organizing extension spike protein, MSN (HGNC:7373)
Calculated MW	78 kDa
Application Details	Western blot, 0.25-0.5 µg/ml, Human, Mouse, Rat, Monkey Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human Flow Cytometry, 1-3 µg/1x^6 cells, Human
Contents	Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clone Names	Clone: 8D4
Immunogen	E.coli-derived human Moesin/MSN recombinant protein (Position: R184-K568).
Purification	Immunogen affinity purified.
Storage	At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Protein Information
Name	MSN (HGNC:7373)
Function	Ezrin-radixin-moesin (ERM) family protein that connects the actin cytoskeleton to the plasma membrane and thereby regulates the structure and function of specific domains of the cell cortex. Tethers actin filaments by oscillating between a resting and an activated state providing transient interactions between moesin and the actin cytoskeleton (PubMed:10212266). Once phosphorylated on its C-terminal threonine, moesin is activated leading to interaction with F-actin and cytoskeletal rearrangement (PubMed:10212266). These rearrangements regulate many cellular processes, including cell shape determination, membrane transport, and signal transduction (PubMed:12387735, PubMed:15039356). The role of moesin is particularly important in immunity acting on both T and B-cells homeostasis and self-tolerance, regulating lymphocyte egress from lymphoid organs (PubMed:9298994, PubMed:9616160). Modulates phagolysosomal biogenesis in macrophages (By similarity). Also participates in immunologic synapse formation (PubMed:27405666).
Cellular Location	Cell membrane; Peripheral membrane protein {ECO:0000250\|UniProtKB:P26041}; Cytoplasmic side {ECO:0000250\|UniProtKB:P26041}. Cytoplasm, cytoskeleton {ECO:0000250\|UniProtKB:P26041}. Apical cell membrane {ECO:0000250\|UniProtKB:P26041}; Peripheral membrane protein {ECO:0000250\|UniProtKB:P26041}; Cytoplasmic side {ECO:0000250\|UniProtKB:P26041}. Cell projection, microvillus membrane {ECO:0000250\|UniProtKB:P26041}; Peripheral membrane protein {ECO:0000250\|UniProtKB:P26041}; Cytoplasmic side {ECO:0000250\|UniProtKB:P26041}. Cell projection, microvillus {ECO:0000250\|UniProtKB:P26041}. Note=Phosphorylated form is enriched in microvilli-like structures at apical membrane. Increased cell membrane localization of both phosphorylated and non-phosphorylated forms seen after thrombin treatment (By similarity). Localizes at the uropods of T lymphoblasts. {ECO:0000250\|UniProtKB:P26041, ECO:0000269\|PubMed:18586956, ECO:0000269\|PubMed:9298994}
Tissue Location	In all tissues and cultured cells studied.

Research Areas

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Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

Background

Moesin is a protein that in humans is encoded by the MSN gene. It is mapped to Xq12. Moesin (for membrane-organizing extension spike protein) is a member of the ERM family which includes ezrin and radixin. ERM proteins appear to function as cross-linkers between plasma membranes and actin-based cytoskeletons. Moesin is localized to filopodia and other membranous protrusions that are important for cell-cell recognition and signaling and for cell movement.

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Cat# ABO15093

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