Anti-Syk antibody [EP573Y] - BSA and Azide free

7 product images

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  Western blot - Anti-Syk antibody [EP573Y] - BSA and Azide free (ab190176)

  False colour image of Western blot: Anti-Syk antibody [EP573Y] 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-Syk antibody [EP573Y] ab40781 was shown to bind specifically to Syk. A band was observed at 70/72 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SYK knockout cell line Human SYK knockout HEK-293T cell line ab282649 (knockout cell lysate Human SYK knockout HEK-293T cell lysate ab283048). To generate this image, wild-type and SYK knockout HEK-293T 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-Syk antibody [EP573Y] (Anti-Syk antibody [EP573Y] ab40781) at 1/1000 dilution

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: SYK knockout HEK-293T cell lysate at 20 µg

  Lane 2: Western blot - Human SYK knockout HEK-293T cell line (Human SYK knockout HEK-293T cell line ab282649)

  Performed under reducing conditions.

  Predicted band size: 72 kDa

  Observed band size: 70 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syk antibody [EP573Y] - BSA and Azide free (ab190176)

  Immunohistochemistry of paraffin embedded Human spleen tissue section labelling Syk with Anti-Syk antibody [EP573Y] ab40781 at 1:5000 dilution (0.44 μg/ml).  Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). A ready to use ImmunoHistoProbe one step HRP Polymer (ready to use) was used as a secondary antibody at 1:0 dilution. PBS instead of primary antibody was used for negative control. Hematoxylin was used as a counterstain.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Syk antibody [EP573Y] ab40781).

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  Western blot - Anti-Syk antibody [EP573Y] - BSA and Azide free (ab190176)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Syk antibody [EP573Y] ab40781).

  All lanes: Western blot - Anti-Syk antibody [EP573Y] (Anti-Syk antibody [EP573Y] ab40781) at 1/1000 dilution

  Lane 1: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg

  Lane 2: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysates at 15 µg

  Lane 3: WEHI-231 (Mouse B cell lymphoma B lymphocyte ) whole cell lysates at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 72 kDa

  Observed band size: 72 kDa

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  Western blot - Anti-Syk antibody [EP573Y] - BSA and Azide free (ab190176)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Syk antibody [EP573Y] ab40781).

  All lanes: Western blot - Anti-Syk antibody [EP573Y] (Anti-Syk antibody [EP573Y] ab40781) at 1/1000 dilution

  All lanes: Human bone marrow lysates at 15 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Predicted band size: 72 kDa

  Observed band size: 72 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Syk antibody [EP573Y] - BSA and Azide free (ab190176)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Syk antibody [EP573Y] ab40781)
  Anti-Syk antibody [EP573Y] ab40781 was shown to react with SYK in wild-type THP-1 cells in immunocytochemistry with loss of signal observed in SYK knockout cell line Human SYK knockout THP-1 cell line ab288700. 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 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with Anti-Syk antibody [EP573Y] ab40781 at 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).
  
  This data was 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.

• 

  Western blot - Anti-Syk antibody [EP573Y] - BSA and Azide free (ab190176)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Syk antibody [EP573Y] ab40781)

  
  Anti-Syk antibody [EP573Y] ab40781 was shown to react with SYK in wild-type THP-1 cells in Western blot with loss of signal observed in SYK knockout cell line Human SYK knockout THP-1 cell line ab288700. Wild-type THP-1 and SYK knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-Syk antibody [EP573Y] ab40781 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.
  
  This data was 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-Syk antibody [EP573Y] (Anti-Syk antibody [EP573Y] ab40781) at 1/1000 dilution

  Lane 1: Wild-type THP-1 lysate at 20 µg

  Lane 2: Western blot - Human SYK knockout THP-1 cell line (Human SYK knockout THP-1 cell line ab288700) at 20 µg

  Observed band size: 72 kDa

• 

  Western blot - Anti-Syk antibody [EP573Y] - BSA and Azide free (ab190176)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Syk antibody [EP573Y] ab40781).
  Western blot: Anti-SYK antibody [EP573Y] (Anti-Syk antibody [EP573Y] ab40781) 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-Syk antibody [EP573Y] ab40781 was shown to bind specifically to SYK. A band was observed at 72 kDa in wild-type THP-1 cell lysates with no signal observed at this size in SYK knockout cell line Human SYK knockout THP-1 cell line ab288700 (knockout cell lysate ab289593). To generate this image, wild-type and SYK knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Syk antibody [EP573Y] (Anti-Syk antibody [EP573Y] ab40781) at 1/1000 dilution

  Lanes 1 - 4: Western blot at 20 µg

  Lane 2: Western blot - Human SYK knockout THP-1 cell line (Human SYK knockout THP-1 cell line ab288700)

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 72 kDa

  Observed band size: 72 kDa