WB
Western blot analysis of Survivin on different lysates with Rabbit anti-Survivin antibody at 1/1,000 dilution. Lane 1: Jurkat cell lysate, Lane 2: HeLa cell lysate, Lane 3: 293T cell lysate, Lane 4: RAW264.7 cell lysate, Lane 5: NIH/3T3 cell lysate, Lane 6: C6 cell lysate, Lane 7: PC-12 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 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 1/1,000 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-Survivin antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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.IHC
Immunohistochemical analysis of paraffin-embedded human lymph node tissue using anti-Survivin antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/400) 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.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling Survivin with Rabbit anti-Survivin 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-Survivin 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 C6 cells labeling Survivin. Cells were fixed and permeabilized. 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).| Product Name | Survivin Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | The baculovirus protein p35 inhibits virally-induced apoptosis of invertebrate and mammalian cells and may function to impair the clearing of virally infected cells by the immune system of the host. This is accomplished at least in part by the ability of p35 to block both TNF- and FAS-mediated apoptosis through the inhibition of the ICE family of serine proteases. Two mammalian homologs of baculovirus p35, referred to as inhibitor of apoptosis protein (IAP) 1 and 2, share an amino-terminal baculovirus IAP repeat (BIR) motif and a carboxy-terminal RING finger. Although the c-IAPs do not directly associate with the TNF receptor (TNF-R), they efficiently block TNF-mediated apoptosis through their interaction with the downstream TNF-R effectors, TRAF1 and TRAF2. Additional IAP family members include ILP (for IAP-like protein) and survivin. ILP inhibits activated caspase-3, leading to the resistance of FAS-mediated apoptosis. Survivin (also designated TIAP) is expressed during the G2/M phase of the cell cycle and associates with microtubules of the mitotic spindle. Increased caspase-3 activity is detected when a disruption of survivin-microtubule interactions occurs. |
| Immunogen | Synthetic peptide within N-terminal human Survivin. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000 IHC:1:200-1:500 ICC/IF:1:100 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | API4 antibody Apoptosis inhibitor 4 antibody Apoptosis inhibitor survivin antibody Apoptosis inhibitor4 antibody Baculoviral IAP repeat containing 5 antibody Baculoviral IAP repeat containing protein 5 antibody Baculoviral IAP repeat-containing protein 5 antibody BIRC 5 antibody BIRC5 antibody BIRC5_HUMAN antibody EPR 1 antibody IAP4 antibody Survivin variant 3 alpha antibody SVV antibody TIAP antibody |
|---|---|
| Molecular Weight(MW) | 16 kDa |
| Cellular Localization | Cytoplasm, Nucleus, Chromosome, Midbody. |
| SwissProt ID | O15392 |
|---|
WB
Western blot analysis of Survivin on different lysates with Rabbit anti-Survivin antibody at 1/1,000 dilution. Lane 1: Jurkat cell lysate, Lane 2: HeLa cell lysate, Lane 3: 293T cell lysate, Lane 4: RAW264.7 cell lysate, Lane 5: NIH/3T3 cell lysate, Lane 6: C6 cell lysate, Lane 7: PC-12 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 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 1/1,000 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-Survivin antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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.
IHC
Immunohistochemical analysis of paraffin-embedded human lymph node tissue using anti-Survivin antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/400) 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.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling Survivin with Rabbit anti-Survivin 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-Survivin 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 C6 cells labeling Survivin. Cells were fixed and permeabilized. 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).| Application Notes | WB:1:1000 IHC:1:200-1:500 ICC/IF:1:100 FC:1:1000 |
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| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
