Anti-Survivin antibody (ab8228)

Immunocytochemistry/ Immunofluorescence - Survivin antibody (ab8228)

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-iT^TM 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).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Survivin antibody (ab8228)

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.

This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Survivin antibody (ab8228)

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.

This image is a courtesy of Anonymous Abreview

Immunocytochemistry/ Immunofluorescence - Survivin antibody (ab8228)

IHC image of Survivin staining in human lymphoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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.

Western blot - Survivin antibody (ab8228)

All lanes : Anti-Survivin antibody (ab8228) at 2 µg/ml

Lane 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.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 16,18 kDa (why is the actual band size different from the predicted?)


Exposure time : 8 minutes

This antibody was blocked using 3% milk. Using a higher concentration of blocking buffer may help to further reduce non-specific bands.