Anti-CD3D antibody [EP4426]

6 product images

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  Flow Cytometry - Anti-CD3D antibody [EP4426] (ab109531)

  Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling CD3D with Purified ab109531 at 1:20 dilution (5 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

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  Western blot - Anti-CD3D antibody [EP4426] (ab109531)

  All lanes: Western blot - Anti-CD3D antibody [EP4426] (ab109531) at 1/1000 dilution

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

  Lane 2: HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate

  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: 19 kDa

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  Immunoprecipitation - Anti-CD3D antibody [EP4426] (ab109531)

  Purified ab109531 at 1/50 dilution (2μg) immunoprecipitating CD3D in Jurkat whole cell lysate.
  Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
  Lane 2 (+): ab109531 + Jurkat whole cell lysate.
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109531 in Jurkat whole cell lysate.
  VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/1000 dilution) was used for Western blotting.
  Blocking Buffer and concentration: 5% NFDM/TBST.
  Diluting buffer and concentration: 5% NFDM/TBST.
  Observed band size: 23 kDa

  All lanes: Immunoprecipitation - Anti-CD3D antibody [EP4426] (ab109531)

  Predicted band size: 19 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-CD3D antibody [EP4426] (ab109531)

  Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD3D with Purified ab109531 at 1:50 dilution (2.6 μg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD3D with Purified ab109531 at 1:100 dilution (1.3 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

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  Western blot - Anti-CD3D antibody [EP4426] (ab109531)

  Lanes 1 - 6: Merged signal (red and green). Green - ab109531 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 ab109531 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) 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 a 1:10000 dilution for 1hr at room temperature and then imaged.

  All lanes: Western blot - Anti-CD3D antibody [EP4426] (ab109531) at 1/1000 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

  Performed under reducing conditions.

  Predicted band size: 19 kDa

  Observed band size: 23 kDa