Bovine serum albumin a2153

OrmoPrime08, S1805 and SU-8 photoresist,
SU-8 2000 thinner, and an AZ303 developer were purchased from Micro
Resist Technology (Germany). Dimethyl sulfoxide (DMSO), acetic acid,
propylene glycol monomethyl ether acetate, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
(EDC) were obtained from Sigma Aldrich (Germany). pNIPAAm-based terpolymer
composed of N-isopropylacrylamide, methacrylic acid,
and 4-methacryloyloxybenzophenone (in a ratio of 94:5:1), benzophenone
disulfide, and 4-sulfotetrafluorophenol (TFPS) were synthesized in
our laboratory as previously reported.^{43 (link)−45 (link)}IgG from mouse
serum (mIgG, I 5381), Tween 20 (P9416), and bovine serum albumin (A2153)
were purchased from Sigma Aldrich (Austria). Phosphate-buffered saline
(PBS) and sodium acetate were obtained from VWR Chemicals (Austria).
Alexa Fluor 790 goat anti-mouse IgG (a-mIgG, A11375) was acquired
from Life Technologies (Eugene, OR).

Immunohistological stains for Ki-67 and AFP were carried out on all malignant and corresponding peri-malignant liver tissues. Anti-Ki 67 (The primary antibodies used were MIB-1 mAb (×50, Immunotech SA) for Ki-67) or anti-AFP antibody “Polyclonal Rabbit Anti-Human Alpha-1-Fetoprotein, code No. A 0008” (DAKO) were used for immunohistochemical detection of the expression of Ki 67 or AFP in tissue.
Tissue sections were processed for IHC analysis as follows: IHC examinations were carried out on 3 µm thick sections. Unmasking was performed with 10 mM sodium citrate buffer, pH 6.0, at 90°C for 30 min. For Sections were incubated in 0.03% hydrogen peroxide for 10 min at room temperature, to remove endogenous peroxidase activity, and then in blocking serum (0.04% bovine serum albumin, A2153, Sigma-Aldrich, Shanghai, China, and 0.5% normal goat serum X0907, in PBS) (Dako Corporation, Carpinteria, CA, USA), for 30 min at room temperature.
The antibody was used at a dilution of 1:100. The antibody was incubated overnight at 4°C. Sections were then washed three times for 5 min in phosphate buffer saline (PBS). Non-specific staining was blocked by 5% normal serum for 30 min at room temperature. Finally, staining was developed with diaminobenzidine substrate and sections were counterstained with hematoxylin. PBS replaced both Ki 67 and AFP antibodies in negative controls.