Anti-Survivin antibody
| Name | Anti-Survivin antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab8228 |
| Prices | $394.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | WB ICC/IF ICC/IF IHC-P IP |
| Species Reactivities | Human, Bovine, Cat, Dog, Pig |
| Antigen | Synthetic peptide derived from within residues 100 to the C-terminus of Human Survivin |
| Blocking Peptide | Human Survivin peptide |
| Description | Rabbit Polyclonal |
| Gene | BIRC5 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
ICC/IF image of ab8228 stained SH-SY-5Y cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab8228, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
ab8228 staining Survivin in human breast cancer tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block then 10 minutes/protein block) for 15 minutes at 20°C. The primary antibody was diluted, 1/250 (or 1/1500) and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.See Abreview
ab8228 staining Survivin in human stomach cancer tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in EDTA for 30 minutes at 1000C. The primary antibody was diluted 1/200 and incubated with sample for 30 minutes at 25°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.See Abreview
IHC image of Survivin staining in human lymphoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8228, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
All lanes : Anti-Survivin antibody (ab8228) at 2 µg/mlLane 1 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate Lane 2 : Kidney (Human) Tissue Lysate - fetal normal tissue (ab30204)Lysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Observed band size : 16,18 kDa (why is the actual band size different from the predicted?)Exposure time : 8 minutesThis antibody was blocked using 3% milk. Using a higher concentration of blocking buffer may help to further reduce non-specific bands.
Survivin was immunoprecipitated using 0.5mg HL60 Whole cell lysate, 5µg of Rabbit polyclonal to Survivin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, HL60 Whole cell lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab8228.Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.Band: 18kDa; Survivin
Product References
Biological function and prognostic significance of peroxisome - Biological function and prognostic significance of peroxisome
Yang L, Zhang H, Zhou ZG, Yan H, Adell G, Sun XF. Clin Cancer Res. 2011 Jun 1;17(11):3760-70.
Combination of sapacitabine and HDAC inhibitors stimulates cell death in AML and - Combination of sapacitabine and HDAC inhibitors stimulates cell death in AML and
Green SR, Choudhary AK, Fleming IN. Br J Cancer. 2010 Oct 26;103(9):1391-9.