Mouse monoclonal pcna antibody

All animal studies were conducted strictly in accordance with the guidelines of the Animal Care and Use Committee of Wuhan University. Four-week-old female BALB/c nude mice were purchased from Beijing HFK Bio-Technology (Beijing, China) and randomly divided into two group of six each. A total of 1 × 10⁷ BxPC-3 cells were resuspended in 100 μL of PBS and subcutaneously injected into the left flank of the mice. Tumor volume was measured with Vernier calipers every 5 days. Tumor volume was calculated as volume (mm³) = [width2 (mm²) × length (mm)]/2. When the tumor reached an average volume of 100 mm³, the tumor-bearing nude mice received an intratumoral multisite injection of antagomir control (5 nmol) or miR-324-5p antagomir (5 nmol) (GenePharma, China) once every 5 days. All mice were sacrificed after 35 days. All tumors were stripped, weighed, and divided either for detection of miR-324-5p expression by quantitative reverse transcription PCR or fixed in formalin for immunohistochemistry (IHC). The mouse PCNA monoclonal antibody (1:100, Santa Cruz Biotechnology, USA) was used to detect PCNA expression in mice tumor tissues. ImageJ software was used to quantitatively analyze and count positive nuclear ratios.