Recombinant Anti-Moesin antibody [EP1863Y] (ab52490)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1863Y] to Moesin
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Moesin antibody [EP1863Y]
See all Moesin primary antibodies -
Description
Rabbit monoclonal [EP1863Y] to Moesin -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
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Positive control
- WB: Hela, Wild-type HAP1, HeLa and Raji whole cell lysate IHC-P: Human tonsil tissue ICC/IF: HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1863Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab52490 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 |
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Flow Cyt (Intra) |
1/30 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (2) |
1/100 - 1/250.
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WB | (3) |
1/20000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
For unpurified use at 1/1000 - 1/10000. |
IP |
1/20 - 1/70.
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IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/30 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/250. |
WB
1/20000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa). For unpurified use at 1/1000 - 1/10000. |
IP
1/20 - 1/70. |
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Probably involved in connections of major cytoskeletal structures to the plasma membrane. -
Tissue specificity
In all tissues and cultured cells studied. -
Sequence similarities
Contains 1 FERM domain. -
Post-translational
modificationsPhosphorylation on Thr-558 is crucial for the formation of microvilli-like structures. -
Cellular localization
Cell membrane. Cytoplasm > cytoskeleton. Apical cell membrane. Cell projection > microvillus membrane. Phosphorylated form is enriched in microvilli-like structures at apical membrane (By similarity). Increased cell membrane localization of both phosphorylated and non-phosphorylated forms seen after thrombin treatment. - Information by UniProt
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Database links
- Entrez Gene: 4478 Human
- Entrez Gene: 17698 Mouse
- Entrez Gene: 81521 Rat
- Omim: 309845 Human
- SwissProt: P26038 Human
- SwissProt: P26041 Mouse
- SwissProt: O35763 Rat
- Unigene: 87752 Human
see all -
Alternative names
- Epididymis luminal protein 70 antibody
- HEL70 antibody
- Membrane organizing extension spike protein antibody
see all
Images
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All lanes : Anti-Moesin antibody [EP1863Y] (ab52490) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MSN knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Moesin antibody [EP1863Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab52490 was shown to bind specifically to Moesin. A band was observed at 75 kDa in wild-type HeLa cell lysates with no signal observed at this size in MSN knockout cell line ab265020 (knockout cell lysate ab257542). To generate this image, wild-type and MSN knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-Moesin antibody [EP1863Y] (ab52490) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Moesin knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : Raji whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 68 kDaLanes 1 - 4: Merged signal (red and green). Green - ab52490 observed at 75 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab52490 was shown to specifically react with Moesin in wild-type HAP1 cells as signal was lost in Moesin knockout cells. Wild-type and Moesin knockout samples were subjected to SDS-PAGE. Ab52490 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Moesin antibody [EP1863Y] (ab52490)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling Moesin with purified ab52490 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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ab52490 was shown to react with MSN in wild-type HeLa cells in Immunocytochemistry with loss of signal observed in MSN knockout cell line ab265020. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1/10000. The cells were then incubated with ab52490 at 1/200 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
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Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Moesin with purified ab52490 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
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Intracellular Flow Cytometry analysis of HeLa cells labelling Moesin with purified ab52490 at 1/30 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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All lanes : Anti-Moesin antibody [EP1863Y] (ab52490) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MSN knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 68 kDaab52490 was shown to react with MSN in wild-type HeLa cells in Western blot with loss of signal observed in MSN knockout cell line ab265020 (MSN knockout cell lysate ab257542). Wild-type HeLa and MSN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab52490 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2ug/mL before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
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ab52490 (purified) at 1/20 immunoprecipitating Moesin in HeLa whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/10,000) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-Moesin antibody [EP1863Y] (ab52490) at 1/20000 dilution (purified)
Lane 1 : HeLa cell lysate
Lane 2 : Raji cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 68 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
Anti-Moesin antibody [EP1863Y] (ab52490) at 1/50000 dilution (purified) + C6 cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 68 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
Overlay histogram showing HeLa cells stained with unpurified ab52490 (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 (ab52490, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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All lanes : Anti-Moesin antibody [EP1863Y] (ab52490) at 1/20000 dilution (purified)
Lane 1 : Neuro-2a cell lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat heart lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 68 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (37)
ab52490 has been referenced in 37 publications.
- Caster DJ et al. Patients with Proliferative Lupus Nephritis Have Autoantibodies That React to Moesin and Demonstrate Increased Glomerular Moesin Expression. J Clin Med 10:N/A (2021). PubMed: 33669337
- Li YQ et al. Moesin as a prognostic indicator of lung adenocarcinoma improves prognosis by enhancing immune lymphocyte infiltration. World J Surg Oncol 19:109 (2021). PubMed: 33838692
- Uysal-Onganer P et al. Peptidylarginine Deiminase Inhibitor Application, Using Cl-Amidine, PAD2, PAD3 and PAD4 Isozyme-Specific Inhibitors in Pancreatic Cancer Cells, Reveals Roles for PAD2 and PAD3 in Cancer Invasion and Modulation of Extracellular Vesicle Signatures. Int J Mol Sci 22:N/A (2021). PubMed: 33573274
- Zhang Y et al. Oocyte-derived microvilli control female fertility by optimizing ovarian follicle selection in mice. Nat Commun 12:2523 (2021). PubMed: 33953177
- Zhang L et al. STK10 knockout inhibits cell migration and promotes cell proliferation via modulating the activity of ERM and p38 MAPK in prostate cancer cells. Exp Ther Med 22:851 (2021). PubMed: 34149897