PRODUCT CODE: ET1701-57

CD3D Recombinant Rabbit Monoclonal Antibody [JJ08-97] (ET1701-57)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

Western blot analysis of CD3D on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-57, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of CD3D on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-57, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • ICC staining of CD3D in Jurkat cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-57, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human spleen 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.
  • Immunohistochemical analysis of paraffin-embedded human kidney 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.
  • 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.
  • Flow cytometric analysis of CD3D was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-57, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of CD3D on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-57, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

CD3D Recombinant Rabbit Monoclonal Antibody [JJ08-97] (ET1701-57)

Immunogen

Recombinant protein within human cd3d aa 106-171.

Host

Rabbit

Positive Control

Jurkat, A431, Hela, SW480, human tonsil tissue, human spleen tissue, human kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JJ08-97

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

19 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

CD3D

SYNONYMS

CD3 antigen delta subunit antibody; CD3 delta antibody; CD3d antibody; CD3d antigen delta polypeptide (TiT3 complex) antibody; CD3d molecule delta (CD3-TCR complex) antibody; CD3D_HUMAN antibody; IMD19 antibody; OKT3 delta chain antibody; T cell receptor T3 delta chain antibody; T-cell receptor T3 delta chain antibody; T-cell surface glycoprotein CD3 delta chain antibody; T3D antibody

TISSUE SPECIFICITY

CD3D is mostly present on T-lymphocytes with its TCR-CD3 partners. Present also in fetal NK-cells.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated on Tyr residues after T-cell receptor triggering by LCK in association with CD4/CD8.

SUBCELLULAR LOCATION

Membrane.

FUNCTION

The T cell antigen receptor (TCR) recognizes foreign antigens and translates such recognition events into intracellular signals that elicit a change in the cell from a dormant to an activated state. Much of this signaling process can be attributed to a multisubunit complex of proteins that associates directly with the TCR. This complex has been designated CD3 (cluster of differentiation 3). It is composed of five invariant polypeptide chains that associate to form three dimers: a heterodimer of gamma and epsilon chains (γε), a heterodimer of delta and epsilon chains (δε) and a homodimer of two zeta chains (ΩΩ) or a heterodimer of zeta and eta chains (Ωh). The Ω and h chains are encoded by the same gene but differ in their carboxyl-terminal ends due to an alternative splicing event. The γ, ε and δ chains each contain a single copy of a conserved immunoreceptor tyrosine-based activation motif (ITAM). In contrast, the Ω chain contains three consecutive copies of the same motif. Phosphorylated ITAMs act as docking sites for protein kinases such as ZAP-70 and Syk and are also capable of regulating their kinase activity. The crystal structure of the ZAP-70 SH2 domains bound to the Ω chain ITAMs has been solved.