WB
Western blot analysis of CD27 on different lysates with Rabbit anti-CD27 antibody at 1/2,000 dilution. Lane 1: Raji cell lysate, Lane 2: Ramos cell lysate, Lane 3: MCF7 cell lysate (negative), Lane 4: Daudi cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 30 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 1/2,000 dilution was used in 5% NFDM/TBST 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 human tonsil tissue with Rabbit anti-CD27 antibody 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 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.ICC/IF
Immunocytochemistry analysis of Raji cells labeling CD27 with Rabbit anti-CD27 antibody 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-CD27 antibody at 1/100 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 human peripheral blood lymphocytes labeling CD27. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (black). 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 at 1/1,000 dilution for 30 minutes at +4℃.IP
CD27 was immunoprecipitated from 0.2 mg Raji cell lysate with Rabbit anti-CD27 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-CD27 antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Raji cell lysate (input), Lane 2: Rabbit anti-CD27 antibody IP in Raji cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-CD27 antibody in Raji cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes.| Product Name | CD27 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human CD27 aa 1-191. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:2000 IHC:1:200 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CD27 |
|---|---|
| Gene Synonyms | T14 S152 Tp55 TNFRSF7 S152. LPFS2 |
| Gene Full Name | CD27 molecule |
| Gene Summary | The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor is required for generation and long-term maintenance of T cell immunity. It binds to ligand CD70, and plays a key role in regulating B-cell activation and immunoglobulin synthesis. This receptor transduces signals that lead to the activation of NF-kappaB and MAPK8/JNK. Adaptor proteins TRAF2 and TRAF5 have been shown to mediate the signaling process of this receptor. CD27-binding protein (SIVA), a proapoptotic protein, can bind to this receptor and is thought to play an important role in the apoptosis induced by this receptor. [provided by RefSeq, Jul 2008] |
| Alternative Names | CD 27 antibody CD27 antibody CD27 antigen antibody CD27 molecule antibody CD27_HUMAN antibody CD27L receptor antibody LPFS2 antibody MGC20393 antibody OTTHUMP00000238557 antibody S152 antibody CD 27 antibody CD27 antibody CD27 antigen antibody CD27 molecule antibody CD27_HUMAN antibody CD27L receptor antibody LPFS2 antibody MGC20393 antibody OTTHUMP00000238557 antibody S152 antibody T cell activation antigen CD27 antibody T cell antivation antigen S152 antibody T-cell activation antigen CD27 antibody T14 antibody TNFRSF 7 antibody TNFRSF7 antibody TNFSF7 antibody Tp 55 antibody Tp55 antibody Tumor necrosis factor receptor superfamily member 7 antibody |
| Molecular Weight(MW) | 29kDa(Observed band size: 55kDa) |
| Cellular Localization | Cell membrane. |
WB
Western blot analysis of CD27 on different lysates with Rabbit anti-CD27 antibody at 1/2,000 dilution. Lane 1: Raji cell lysate, Lane 2: Ramos cell lysate, Lane 3: MCF7 cell lysate (negative), Lane 4: Daudi cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 30 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 1/2,000 dilution was used in 5% NFDM/TBST 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 human tonsil tissue with Rabbit anti-CD27 antibody 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 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.
ICC/IF
Immunocytochemistry analysis of Raji cells labeling CD27 with Rabbit anti-CD27 antibody 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-CD27 antibody at 1/100 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 human peripheral blood lymphocytes labeling CD27. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (black). 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 at 1/1,000 dilution for 30 minutes at +4℃.
IP
CD27 was immunoprecipitated from 0.2 mg Raji cell lysate with Rabbit anti-CD27 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-CD27 antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Raji cell lysate (input), Lane 2: Rabbit anti-CD27 antibody IP in Raji cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-CD27 antibody in Raji cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes.| Application Notes | WB:1:2000 IHC:1:200 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
