Anti-CD3 epsilon antibody [SP7], prediluted

4 product images

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  Flow Cytometry (Intracellular) - Anti-CD3 epsilon antibody [SP7], prediluted (ab21703)

  This image was generated using a previous batch manufactured using hybridoma production method.

  Human peripheral blood lymphocytes stained with Anti-CD3 epsilon antibody [SP7] ab16669 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (Anti-CD3 epsilon antibody [SP7] ab16669, 1/1000 dilution) for 30 min at 4°C. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.

  This data was developed using the undiluted version of this antibody (Anti-CD3 epsilon antibody [SP7] ab16669).

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  Western blot - Anti-CD3 epsilon antibody [SP7], prediluted (ab21703)

  This image was generated using a previous batch manufactured using hybridoma production method.

  Lanes 1 - 6: Merged signal (red and green). Green – Anti-CD3 epsilon antibody [SP7] ab16669 observed at 23 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

  This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with Anti-CD3 epsilon antibody [SP7] ab16669 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  This data was developed using the undiluted version of this antibody (Anti-CD3 epsilon antibody [SP7] ab16669).

  All lanes: Western blot - Anti-CD3 epsilon antibody [SP7], prediluted (ab21703) at 1/25 dilution

  Lane 1: THP1 whole cell lysate (-ve control) at 15 µg

  Lane 2: Raji whole cell lysate (-ve control) at 15 µg

  Lane 3: Jurkat whole cell lysate at 15 µg

  Lane 4: Human Thymus tissue lysate at 15 µg

  Lane 5: Mouse Thymus tissue lysate at 15 µg

  Lane 6: Rat Thymus tissue lysate at 15 µg

  Secondary

  All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

  Predicted band size: 19 kDa

  Observed band size: 23 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [SP7], prediluted (ab21703)

  Immunohistochemical analysis of paraffin-embedded Human tonsil labeling CD3 epsilon with ab21703 at 1/150 (7 μg/ml). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab21703 for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. DAB was used as the chromogen. Counterstained with Hematoxylin and mounted with DPX.

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  Western blot - Anti-CD3 epsilon antibody [SP7], prediluted (ab21703)

  This data was developed using Anti-CD3 epsilon antibody [SP7] ab16669, the same antibody clone in a different buffer formulation.
  

  Negative control: U-937, Raji, brain.

  All lanes: Western blot - Anti-CD3 epsilon antibody [SP7] (Anti-CD3 epsilon antibody [SP7] ab16669) at 1/1000 dilution

  Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

  Lane 2: MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

  Lane 3: U-937 (Human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

  Lane 4: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

  Lane 5: EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

  Lane 6: Mouse thymus lysate at 20 µg

  Lane 7: Mouse brain lysate at 20 µg

  Lane 8: Rat thymus lysate at 20 µg

  Lane 9: Rat brain lysate at 20 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

  Observed band size: 23 kDa

  Exposure time: 180s