CD3D Recombinant Rabbit Monoclonal Antibody [JJ08-97]
cat.: ET1701-57
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: JJ08-97
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 19 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human CD3D aa 60-171 / 171.
Positive control: Jurkat cell lysates, Jurkat, human spleen tissue, human thymus tissue, human tonsil tissue.
Subcellular location: Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:1,000
1:100
1:200-1:1,000
1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: P04234 Human
Alternative names: CD3 antigen delta subunit CD3 delta CD3d CD3d antigen delta polypeptide (TiT3 complex) CD3d molecule delta (CD3-TCR complex) CD3D_HUMAN IMD19 OKT3 delta chain T cell receptor T3 delta chain T-cell receptor T3 delta chain T-cell surface glycoprotein CD3 delta chain T3D
Images
ET1701-57_1.jpg Fig1: Western blot analysis of CD3D on Jurkat cell lysates with Rabbit anti-CD3D antibody (ET1701-57) at 1/1,000 dilution.

Lysates/proteins at 15 µg/Lane.

Predicted band size: 19 kDa
Observed band size: 23 kDa

Exposure time: 17 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-57) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1701-57_2.jpg Fig2: Immunocytochemistry analysis of Jurkat cells labeling CD3D with Rabbit anti-CD3D antibody (ET1701-57) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD3D antibody (ET1701-57) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1701-57_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD3D antibody (ET1701-57) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-57) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-57_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human thymus tissue with Rabbit anti-CD3D antibody (ET1701-57) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-57) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-57_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD3D antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-57, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-57_6.jpg Fig6: Flow cytometric analysis of Jurkat cells labeling CD3D.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-57, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.