Anti-Moesin antibody [EP1863Y] - BSA and Azide free

9 product images

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  Immunoprecipitation - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  Anti-Moesin antibody [EP1863Y] 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 (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/10,000) was used for detection. A rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used intead of Anti-Rab11A antibody [EPR7587(B)] ab128913 as a negative control (Lane 3).
  

  Blocking buffer and concentration: 5% NFDM/TBST.

  Diluting buffer and concentration: 5% NFDM /TBST.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Moesin antibody [EP1863Y] ab52490).

  All lanes: Immunoprecipitation - Anti-Moesin antibody [EP1863Y] (Anti-Moesin antibody [EP1863Y] ab52490)

  Predicted band size: 68 kDa

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  Western blot - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Moesin antibody [EP1863Y] ab52490).
  

  False colour image of Western blot: Anti-Moesin antibody [EP1863Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Moesin antibody [EP1863Y] 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 Human MSN (Moesin) knockout HeLa cell line ab265020 (knockout cell lysate Human MSN (Moesin) knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

  All lanes: Western blot - Anti-Moesin antibody [EP1863Y] (Anti-Moesin antibody [EP1863Y] ab52490) 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 (Human MSN (Moesin) knockout HeLa cell line ab265020)

  Performed under reducing conditions.

  Predicted band size: 68 kDa

  Observed band size: 75 kDa

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  Western blot - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  Lanes 1 - 4: Merged signal (red and green). Green - Anti-Moesin antibody [EP1863Y] ab52490 observed at 75 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
  

  Anti-Moesin antibody [EP1863Y] 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. Anti-Moesin antibody [EP1863Y] ab52490 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1,000 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Moesin antibody [EP1863Y] ab52490).

  All lanes: Western blot - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580) at 1/1000 dilution

  Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

  Lane 2: Moesin knockout HAP1 whole cell lysate at 20 µg

  Lane 3: HeLa whole cell lysate at 20 µg

  Lane 4: Raji whole cell lysate at 20 µg

  Predicted band size: 68 kDa

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  Flow Cytometry (Intracellular) - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  Intracellular Flow Cytometry analysis of HeLa cells labelling Moesin with purified Anti-Moesin antibody [EP1863Y] 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.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Moesin antibody [EP1863Y] ab52490).

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  Immunocytochemistry - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Moesin antibody [EP1863Y] ab52490). Anti-Moesin antibody [EP1863Y] ab52490 was shown to react with MSN in wild-type HeLa cells in Immunocytochemistry with loss of signal observed in MSN knockout cell line Human MSN (Moesin) knockout HeLa 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 Anti-Moesin antibody [EP1863Y] 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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  Western blot - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  This data was developed using the same antibody in a different buffer formulation (Anti-Moesin antibody [EP1863Y] ab52490).
  

  Anti-Moesin antibody [EP1863Y] 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 Human MSN (Moesin) knockout HeLa cell line ab265020 (MSN knockout cell lysate Human MSN (Moesin) knockout HeLa 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 Anti-Moesin antibody [EP1863Y] 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] (Anti-Moesin antibody [EP1863Y] ab52490) 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 (Human MSN (Moesin) knockout HeLa cell line ab265020)

  Performed under reducing conditions.

  Predicted band size: 68 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Moesin with purified Anti-Moesin antibody [EP1863Y] ab52490 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

  Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

  Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Moesin antibody [EP1863Y] ab52490).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling Moesin with purified Anti-Moesin antibody [EP1863Y] ab52490 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) 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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Moesin antibody [EP1863Y] ab52490).

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  Flow Cytometry (Intracellular) - Anti-Moesin antibody [EP1863Y] - BSA and Azide free (ab232580)

  Overlay histogram showing HeLa cells stained with unpurified Anti-Moesin antibody [EP1863Y] 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 (Anti-Moesin antibody [EP1863Y] ab52490, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Moesin antibody [EP1863Y] ab52490).