WB
Western blot analysis of CD3 on different lysates with Rabbit anti-CD3 antibody at 1/20,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 4 seconds; 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 at 1/20,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-CD3 antibody at 1/5,000 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 at 1/5,000 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.ICC/IF
Immunocytochemistry analysis of Jurkat (positive) and THP-1 (negative) labeling CD3 with Rabbit anti-CD3 antibody at 1/500 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-CD3 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.ICC/IF
Immunocytochemistry analysis of mouse splenocytes labeling CD3 with Rabbit anti-CD3 antibody at 1/200 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-CD3 antibody at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of MOLT-4 cells labeling CD3. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IF-F
Application: IHC-Fr. Species: Mouse. Site: spleen. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.IF-F
Application: IHC-Fr. Species: Rat. Site: spleen. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.IF-P
Application: IF-Tissue. Species: Mouse. Site: spleen. Sample: Paraffin-embedded section. Antibody concentration: 1/500.IF-P
Application: IF-Tissue. Species: Rat. Site: spleen. Sample: Paraffin-embedded section. Antibody concentration: 1/500.IP
CD3 was immunoprecipitated from 0.2 mg MOLT-4 cell lysate with CD3 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using CD3 Recombinant Rabbit Monoclonal Antibody at 1/10,000 dilution. Alpaca anti-Rabbit IgG Fc secondary antibody at 1/50,000 dilution was used for 1 hour at room temperature. Lane 1: MOLT-4 cell lysate (input). Lane 2: CD3 Recombinant Rabbit Monoclonal Antibody IP in MOLT-4 cell lysate. Lane 3: Rabbit IgG instead of CD3 Recombinant Rabbit Monoclonal Antibody in MOLT-4 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 7 seconds.| Product Name | CD3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within mouse CD3E aa 1-207. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIF-FIF-PIHCIPWB |
| WB:1:5000-1:20000 IHC:1:5000 ICC/IF:1:200-1:2500 FC:1:1000 IF-F:1:500 IF-P:1:500 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CD3E |
|---|---|
| Gene Synonyms | T3E TCRE IMD18 CD3epsilon |
| Gene Full Name | CD3 epsilon subunit of T-cell receptor complex |
| Gene Summary | The protein encoded by this gene is the CD3-epsilon polypeptide, which together with CD3-gamma, -delta and -zeta, and the T-cell receptor alpha/beta and gamma/delta heterodimers, forms the T-cell receptor-CD3 complex. This complex plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. The genes encoding the epsilon, gamma and delta polypeptides are located in the same cluster on chromosome 11. The epsilon polypeptide plays an essential role in T-cell development. Defects in this gene cause immunodeficiency. This gene has also been linked to a susceptibility to type I diabetes in women. [provided by RefSeq, Jul 2008] |
| Alternative Names | T-cell surface glycoprotein CD3 epsilon chain T-cell surface antigen T3/Leu-4 epsilon chain CD3e CD3E T3E |
| Molecular Weight(MW) | 23 kDa(Observed band size: 20 kDa) |
| Cellular Localization | Cell membrane. |
WB
Western blot analysis of CD3 on different lysates with Rabbit anti-CD3 antibody at 1/20,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 4 seconds; 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 at 1/20,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-CD3 antibody at 1/5,000 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 at 1/5,000 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.
ICC/IF
Immunocytochemistry analysis of Jurkat (positive) and THP-1 (negative) labeling CD3 with Rabbit anti-CD3 antibody at 1/500 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-CD3 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
ICC/IF
Immunocytochemistry analysis of mouse splenocytes labeling CD3 with Rabbit anti-CD3 antibody at 1/200 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-CD3 antibody at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of MOLT-4 cells labeling CD3. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IF-F
Application: IHC-Fr. Species: Mouse. Site: spleen. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.
IF-F
Application: IHC-Fr. Species: Rat. Site: spleen. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.
IF-P
Application: IF-Tissue. Species: Mouse. Site: spleen. Sample: Paraffin-embedded section. Antibody concentration: 1/500.
IF-P
Application: IF-Tissue. Species: Rat. Site: spleen. Sample: Paraffin-embedded section. Antibody concentration: 1/500.
IP
CD3 was immunoprecipitated from 0.2 mg MOLT-4 cell lysate with CD3 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using CD3 Recombinant Rabbit Monoclonal Antibody at 1/10,000 dilution. Alpaca anti-Rabbit IgG Fc secondary antibody at 1/50,000 dilution was used for 1 hour at room temperature. Lane 1: MOLT-4 cell lysate (input). Lane 2: CD3 Recombinant Rabbit Monoclonal Antibody IP in MOLT-4 cell lysate. Lane 3: Rabbit IgG instead of CD3 Recombinant Rabbit Monoclonal Antibody in MOLT-4 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 7 seconds.| Application Notes | WB:1:5000-1:20000 IHC:1:5000 ICC/IF:1:200-1:2500 FC:1:1000 IF-F:1:500 IF-P:1:500 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
