ab2098 diluted 1:500 to detect Cyclin T1 in human skin tissue. Tissue was formalin fixed and paraffin embedded. No pre-treatment of sample was required. The image shows the localization of antibody as the precipitated red signal, with a hematoxylin purple nuclear counter stain.

ab2098 (2µg/ml) staining Cyclin T1 in human lymph node using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

Anti-Cyclin T1 antibody (ab2098) at 1/10000 dilution + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 81 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa, 48 kDa, 78 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 16 minutes


The 90 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Cyclin T1.