AB265020

Human MSN (Moesin) knockout HeLa cell line

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Human MSN (Moesin) knockout HeLa cell line available to order. Recommended control: Human wild-type HeLa cell line (ab255448).

View Alternative Names

Epididymis luminal protein 70, HEL70, MOES_HUMAN, MSN, MSN/ALK fusion, Membrane-organizing extension spike protein, Moesin, Moesin/anaplastic lymphoma kinase fusion protein

11 Images

Collaborator

Immunocytochemistry - Human MSN (Moesin) knockout HeLa cell line (AB265020)

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 worked 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.

Lab

Western blot - Human MSN (Moesin) knockout HeLa cell line (AB265020)

False colour image of Western blot : Anti-Moesin antibody [MSN/492] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab216033 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-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-Moesin antibody [MSN/492] (<a href='/en-us/products/primary-antibodies/moesin-antibody-msn-492-ab216033'>ab216033</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MSN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MSN (Moesin) knockout HeLa cell line (ab265020)

Predicted band size: 68 kDa

Observed band size: 75 kDa

false

Lab

Western blot - Human MSN (Moesin) knockout HeLa cell line (AB265020)

False colour image of Western blot : Anti-Moesin antibody [EPR2429(2)] staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab169789 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.

All lanes:

Western blot - Anti-Moesin antibody [EPR2429(2)] (<a href='/en-us/products/primary-antibodies/moesin-antibody-epr24292-ab169789'>ab169789</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MSN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MSN (Moesin) knockout HeLa cell line (ab265020)

Predicted band size: 68 kDa

Observed band size: 75 kDa

false

Collaborator

Western blot - Human MSN (Moesin) knockout HeLa cell line (AB265020)

ab52490 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.

All lanes:

Western blot - Anti-Moesin antibody [EP1863Y] (<a href='/en-us/products/primary-antibodies/moesin-antibody-ep1863y-ab52490'>ab52490</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MSN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MSN (Moesin) knockout HeLa cell line (ab265020)

Predicted band size: 68 kDa

false

Lab

Western blot - Human MSN (Moesin) knockout HeLa cell line (AB265020)

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.

All lanes:

Western blot - Anti-Moesin antibody [EP1863Y] (<a href='/en-us/products/primary-antibodies/moesin-antibody-ep1863y-ab52490'>ab52490</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MSN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MSN (Moesin) knockout HeLa cell line (ab265020)

Predicted band size: 68 kDa

Observed band size: 75 kDa

false

Collaborator

Western blot - Human MSN (Moesin) knockout HeLa cell line (AB265020)

ab169789 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 ab169789 overnight at 4 °C at a 1/10000 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.

All lanes:

Western blot - Anti-Moesin antibody [EPR2429(2)] (<a href='/en-us/products/primary-antibodies/moesin-antibody-epr24292-ab169789'>ab169789</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MSN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MSN (Moesin) knockout HeLa cell line (ab265020)

Predicted band size: 68 kDa

false

Lab

Western blot - Human MSN (Moesin) knockout HeLa cell line (AB265020)

False colour image of Western blot : Anti-Moesin antibody [EPR2428(2)] staining at 2 µg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab151542 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.

All lanes:

Western blot - Anti-Moesin antibody [EPR2428(2)] (<a href='/en-us/products/primary-antibodies/moesin-antibody-epr24282-ab151542'>ab151542</a>) at 2 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MSN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MSN (Moesin) knockout HeLa cell line (ab265020)

Predicted band size: 68 kDa

Observed band size: 75 kDa

false

Immunocytochemistry/ Immunofluorescence - Human MSN (Moesin) knockout HeLa cell line (AB265020)

ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Human MSN (Moesin) knockout HeLa cell line (AB265020)

ab151542 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 ab151542 at 1/200; signal ok 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.

ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Human MSN (Moesin) knockout HeLa cell line (AB265020)

ab169789 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 ab169789 at 1/100; signal ok 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.

Collaborator

Western blot - Human MSN (Moesin) knockout HeLa cell line (AB265020)

ab151542 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 ab151542 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.

All lanes:

Western blot - Anti-Moesin antibody [EPR2428(2)] (<a href='/en-us/products/primary-antibodies/moesin-antibody-epr24282-ab151542'>ab151542</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MSN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MSN (Moesin) knockout HeLa cell line (ab265020)

Predicted band size: 68 kDa

false

Sanger Sequencing - Human MSN (Moesin) knockout HeLa cell line (AB265020)

Sanger seq

Unknown

Sanger Sequencing - Human MSN (Moesin) knockout HeLa cell line (AB265020)

Homozygous : 1 bp insertion in exon 5.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Immunocytochemistry,Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "ICC": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab265020-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab265020 Human MSN (Moesin) knockout HeLa cell line", "number":"AB265020-CMP01", "productcode":"" } ] } } }

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Properties and storage information

Gene name

MSN

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Immunocytochemistry, Sanger Sequencing, Western blot

Zygosity

Homozygous

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO₂. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10⁴ cells/cm². Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines

All seeding densities should be based on cell counts gained by established methods.
A guide seeding density of 2x10⁴ cells/cm² is recommended.
Cells should be passaged when they have achieved 80-90% confluence.

Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Moesin also known as MSN or moesin protein is part of the ERM (ezrin radixin moesin) family which links the actin cytoskeleton to the plasma membrane. It weighs around 68 kDa. This protein is widely expressed in various tissues notably in endothelial cells and other cells with high cytoskeletal dynamics. Moesin plays an essential role in maintaining cell shape polarity and enabling cellular processes like migration and adhesion.

Biological function summary

This protein serves as a crosslinker between the plasma membrane and the actin cytoskeleton influencing signal transduction pathways. Moesin participates significantly in cellular processes like cytokinesis and microvilli formation by binding actin filaments. It is part of several functional complexes ensuring proper cytoskeletal organization and cellular dynamics. Moesin's interactions with other proteins like Rho GTPases regulate its functions aiding cellular morphology and polarity.

Pathways

Moesin is actively involved in the RhoA-ROCK pathway. This pathway is fundamental to actin cytoskeletal reorganization and cell contraction. Moesin interacts with proteins such as radixin and ezrin ensuring cohesion in cytoskeletal rearrangements. Through these interactions Moesin contributes to cell motility and various signaling cascades necessary for cellular responses.

Moesin is associated with conditions such as cancer and immune system disorders. Aberrant expression of moesin can contribute to enhanced cellular migration and invasion which are mechanisms seen in cancer metastasis. Moesin's interaction with proteins like ezrin in cancer progression has been documented. Also in immune disorders altered moesin activity can affect leukocyte transmigration disrupting efficient immune responses.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com