WB
Western blot analysis of CD3 on Jurkat cell lysates with Rabbit anti-CD3 antibody at 0.5μg/ml dilution. Lysates/proteins at 10 µg/Lane. Predicted band size: 23 kDa Observed band size: 23 kDa Exposure time: 2 minutes; 15% 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 0.5μg/ml dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-CD3 antibody at 1/600 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 1/600 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 cells labeling CD3 with Rabbit anti-CD3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.IF-P
Immunofluorescence analysis of paraffin-embedded human lymph nodes tissue labeling CD3 with Rabbit anti-CD3 antibody at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).IP
CD3 was immunoprecipitated from 1mg/ml Jurkat whole cell lysate with at 2ug/ml dilution. Western blot was performed from the immunoprecipitate using at 1ug/ml dilution. Goat anti-Rabbit IgG-HRP antibody was used as the secondary antibody at 1/300,000 dilution. Lane 1: Jurkat whole cell lysate 5 μg (Input). Lane 2: CD3 antibody IP in Jurkat whole cell lysate. Lane 3: Rabbit monoclonal IgG instead of CD3 antibody in Jurkat whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 30 seconds.| Product Name | CD3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within human CD3E aa 158-207/207. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIF-PIHCIPWB |
| WB:1:1000-1:2000 IHC:1:600-1:1000 ICC:1:100 IF-P:1:100-1:200 |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CD247 |
|---|---|
| Gene Synonyms | T3Z CD3H CD3Q CD3Z TCRZ IMD25 CD3ZETA CD3-ZETA |
| Gene Full Name | CD247 molecule |
| Gene Summary | The protein encoded by this gene is T-cell receptor zeta, which together with T-cell receptor alpha/beta and gamma/delta heterodimers, and with CD3-gamma, -delta and -epsilon, forms the T-cell receptor-CD3 complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. Low expression of the antigen results in impaired immune response. Two alternatively spliced transcript variants encoding distinct isoforms have been found for this gene. [provided by RefSeq, Jul 2008] |
| Alternative Names | 4930549J05Rik antibody A430104F18Rik antibody AW552088 antibody Cd247 antibody CD247 antigen antibody CD247 antigen zeta subunit antibody CD247 molecule antibody CD3 antibody CD3 antigen delta subunit antibody CD3 delta antibody 4930549J05Rik antibody A430104F18Rik antibody AW552088 antibody Cd247 antibody CD247 antigen antibody CD247 antigen zeta subunit antibody CD247 molecule antibody CD3 antibody CD3 antigen delta subunit antibody CD3 delta antibody CD3 epsilon antibody CD3 eta antibody CD3 gamma antibody CD3 molecule delta polypeptide antibody CD3 molecule epsilon polypeptide antibody CD3 molecule gamma polypeptide antibody CD3 zeta antibody CD3-DELTA antibody CD3d antibody CD3D antigen delta polypeptide antibody CD3d antigen delta polypeptide (TiT3 complex) antibody CD3d molecule delta antibody CD3d molecule delta CD3 TCR complex antibody CD3d molecule delta (CD3-TCR complex) antibody CD3D_HUMAN antibody CD3E antibody CD3e antigen antibody CD3E antigen epsilon polypeptide antibody CD3e antigen epsilon polypeptide (TiT3 complex) antibody CD3e molecule epsilon antibody CD3e molecule epsilon CD3 TCR complex antibody CD3e molecule epsilon (CD3-TCR complex) antibody CD3epsilon antibody CD3G antibody CD3g antigen antibody CD3G antigen gamma polypeptide antibody CD3g antigen gamma polypeptide (TiT3 complex) antibody CD3g molecule gamma antibody CD3g molecule gamma CD3 TCR complex antibody CD3g molecule gamma (CD3-TCR complex) antibody CD3H antibody CD3Q antibody CD3Z antibody CD3zeta antibody Ctg3 antibody FLJ17620 antibody FLJ17664 antibody FLJ18683 antibody FLJ79544 antibody FLJ94613 antibody IMD19 antibody Leu-4 antibody MGC138597 antibody OKT3 delta chain antibody OTTHUMP00000032544 antibody T cell receptor antibody T cell receptor T3 delta chain antibody T cell receptor T3 gamma chain antibody T cell receptor T3 zeta chain antibody T cell receptor zeta chain antibody T cell surface antigen T3/Leu 4 epsilon chain antibody T cell surface glycoprotein CD3 antibody T cell surface glycoprotein CD3 delta chain antibody T cell surface glycoprotein CD3 epsilon chain antibody T cell surface glycoprotein CD3 gamma chain antibody T cell surface glycoprotein CD3 zeta chain antibody T-cell antigen receptor complex delta subunit of T3 antibody T-cell antigen receptor complex epsilon subunit of T3 antibody T-cell antigen receptor complex gamma subunit of T3 antibody T-cell antigen receptor complex zeta subunit of CD3 antibody T-cell receptor T3 delta chain antibody T-cell receptor T3 gamma chain antibody T-cell surface antigen T3/Leu-4 epsilon chain antibody T-cell surface glycoprotein CD3 delta chain antibody T-cell surface glycoprotein CD3 epsilon chain antibody T-cell surface glycoprotein CD3 gamma chain antibody T3 antibody T3d antibody T3e antibody T3g antibody T3z antibody TCRE antibody TCRk antibody Tcrz antibody TCRzeta antibody |
| Molecular Weight(MW) | 23kDa |
| Cellular Localization | Cell membrane. |
WB
Western blot analysis of CD3 on Jurkat cell lysates with Rabbit anti-CD3 antibody at 0.5μg/ml dilution. Lysates/proteins at 10 µg/Lane. Predicted band size: 23 kDa Observed band size: 23 kDa Exposure time: 2 minutes; 15% 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 0.5μg/ml dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-CD3 antibody at 1/600 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 1/600 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 cells labeling CD3 with Rabbit anti-CD3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
IF-P
Immunofluorescence analysis of paraffin-embedded human lymph nodes tissue labeling CD3 with Rabbit anti-CD3 antibody at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
IP
CD3 was immunoprecipitated from 1mg/ml Jurkat whole cell lysate with at 2ug/ml dilution. Western blot was performed from the immunoprecipitate using at 1ug/ml dilution. Goat anti-Rabbit IgG-HRP antibody was used as the secondary antibody at 1/300,000 dilution. Lane 1: Jurkat whole cell lysate 5 μg (Input). Lane 2: CD3 antibody IP in Jurkat whole cell lysate. Lane 3: Rabbit monoclonal IgG instead of CD3 antibody in Jurkat whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 30 seconds.| Application Notes | WB:1:1000-1:2000 IHC:1:600-1:1000 ICC:1:100 IF-P:1:100-1:200 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
