All lanes : Anti-Survivin antibody [EPR2675] (ab134170) at 1/500 dilution

Lane 1 : HeLa cell lysate
Lane 2 : MOLT4 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Developed using the ECL technique.

Predicted band size: 16 kDa

Secondary antibody - anti-rabbit HRP (ab6721)

All lanes : Anti-Survivin antibody [EPR2675] (ab134170) at 1/300 dilution

Lane 1 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 16 kDa


Exposure time: 1 minute

Blocking and diluting buffer: 5% NFDM/TBST.

Ab134170 staining Survivin in paraffin-embedded rat spleen tissue sections by Immunohistochemistry (IHC-P-paraformaldehyde-fixed, paraffin embedded sections). Samples are incubated in primary antibody at 1:300 dilution (3µg/ml). A Goat Anti-rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1:500 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Hematoxylin was used as a counterstain. Sporadically nuclear staining on rat spleen. 

Ab134170 staining Survivin in C6 (rat glial tumor glial cell) cells by Immunocytochemistry. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:500 dilution (2.5µg/ml). An AlexaFluor^®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution (2µg/ml). Cells were counterstained with an Alexa Fluor^® 594 Anti- Alpha Tubulin [DM1A]- Microtubule Marker at 1:200 dilution (2.5µg/ml). DAPI was used as a nuclear counterstain. Confocal image showing strong midbody (arrow) staining in C6 cell line.

Ab134170 staining Survivin in paraffin-embedded mouse stomach tissue sections by Immunohistochemistry (IHC-P-paraformaldehyde-fixed, paraffin embedded sections). Samples are incubated in primary antibody at 1:300 dilution (3µg/ml). A Goat Anti-rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1:500 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Hematoxylin was used as a counterstain. Sporadically nuclear staining on mouse stomach.

Ab134170 staining Survivin in MCF7 (human breast adenocarcinoma epithelial cell line) cells by Immunocytochemistry. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:500 dilution (2.5µg/ml). An AlexaFluor^®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution (2µg/ml). Cells were counterstained with an Alexa Fluor^® 594 Anti- Alpha Tubulin [DM1A]- Microtubule Marker at 1:200 dilution (2.5µg/ml). DAPI was used as a nuclear counterstain. Confocal image showing strong midbody (arrow) staining in MCF7 cell line. 

Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue, staining Survivin using ab134170 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.