Anti-Moesin antibody [EPR2429(2)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Moesin antibody [EPR2429(2)] (AB169789)

Immunohistochemical analysis of paraffin-embedded, formalin-fixed Human lung tissue, labeling Moesin using ab169789 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Moesin antibody [EPR2429(2)] (AB169789)

Immunohistochemical analysis of paraffin-embedded, formalin-fixed Human stomach tissue, labeling Moesin using ab169789 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Moesin antibody [EPR2429(2)] (AB169789)

Intracellular flow cytometric analysis of permeabilized Raji cells labeling ab169789 (red) at a 1/10 dilution, and negative control cells probed with a rabbit IgG (green).

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Moesin antibody [EPR2429(2)] (AB169789)

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 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.

• IP

Collaborator

Immunoprecipitation - Anti-Moesin antibody [EPR2429(2)] (AB169789)

Immunoprecipitation of MSN in HeLa cells. Lysates were prepared and immunoprecipitation was performed using 1.0 μg of ab169789 pre-coupled to prot.A-Sepharose beads. Samples were washed and processed for western blot with ab169789 at 1/10000. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitate. 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:

Immunoprecipitation - Anti-Moesin antibody [EPR2429(2)] (ab169789)

Predicted band size: 68 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-Moesin antibody [EPR2429(2)] (AB169789)

Immunoprecipitation analysis of pellet from HeLa cell lysate, using anti-Moesin ab169789 at a 1/10 dilution.

All lanes:

Immunoprecipitation - Anti-Moesin antibody [EPR2429(2)] (ab169789)

Predicted band size: 68 kDa

false

• WB

Collaborator

Western blot - Anti-Moesin antibody [EPR2429(2)] (AB169789)

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)] (ab169789) 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 (<a href='/en-us/products/cell-lines/human-msn-moesin-knockout-hela-cell-line-ab265020'>ab265020</a>)

Predicted band size: 68 kDa

false

• WB

Lab

Western blot - Anti-Moesin antibody [EPR2429(2)] (AB169789)

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)] (ab169789) 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 (<a href='/en-us/products/cell-lines/human-msn-moesin-knockout-hela-cell-line-ab265020'>ab265020</a>)

Predicted band size: 68 kDa

Observed band size: 75 kDa

false

• WB

Unknown

Western blot - Anti-Moesin antibody [EPR2429(2)] (AB169789)

All lanes:

Western blot - Anti-Moesin antibody [EPR2429(2)] (ab169789) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Raji cell lysate at 10 µg

Lane 3:

293T cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

standard HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 68 kDa

true