Anti her2 antibody

Cells were fixed with 4% paraformaldehyde and then incubated in immunol staining blocking buffer (1× PBS/5% BSA/0.3% Triton X‐100) for 2 hours to permeabilize the cells and block non‐specific protein–protein interactions. Cells were then incubated with anti‐galectin‐3 antibody (Abcam) or anti‐HER2 antibody (Abcam) followed by Alexa Fluor 594‐secondary antibody (Proteintech). Nuclei were detected using 4′,6‐diamidino‐2‐phenylindole (DAPI) staining. Cells were then examined using a fluorescence microscope.

At the end of the protocol (day 8), mice were killed, and tumor samples were excised, fixed in formalin solution, embedded in paraffin, and then sliced into 5-μm sections using a microtome. Samples were stained by hematoxylin and eosin to visualize the tumor necrosis within different groups. The anti-murine caspase-3 p11 antibody (Santa Cruz Biotechnology, Inc) was used for histochemistry to detect the cell apoptosis in tumors (Figure 5A).
The immunoreaction for caspase-3 and Her-2 (anti–Her-2 antibody; Abcam) in tumor cells was determined by two pathologists (P.Y. and C.R.F.), and the consensus was reached for the final diagnosis. The scores and percentage of tumor cells stained are described as follows [5] (link), [6] (link): no positive cells (−), 1% to 10% of the cells stained (+), 11% to 50% of cells stained (++), and 51% to 100% of the cells stained (+++). We then calculated the percentage of number of mice with positive caspase-3 and Her-2 expression in each group and described them by bars. Comparison with the average mean and peak NB intensities analyzed by the software after the treatment was carried out to find the correlation between NB intensities and IHC results.

Abraxane^® (Albumin Bound, Lot: 6109342) was from Celgene (LLC Melrose Park, IL 60160, USA). Taxol^® (Paclitaxel Injection) was from Bristol-Myers Squibb (Corden Phama Latina S.P.A Via Del Murillo Km 2.800, Sermoneta, Latina, Italy). Anti-HER2 antibody (Lot: ab2428) was from Abcam. DMEM/High Glucose (Cat: SH30022.01), Pen Strep (Penicillin Streptomycin, Lot: 1665735), 0.25% Trypsin-EDTA, and PBS (Phosphate Buffered Saline, Lot: AAL211089) were from HyClone (GE Healthcare Life Sciences). FBS (Fetal Bovine Serum, Lot: 1698221) was from Gibco (Carlsbad, CA, USA). EDC (C₈H₁₇N₃HCL, Cat: 25952-53-8), NHS (C₄H₅NO₂, Cat: 6066-82-6) and NIR-797 isothiocyanate (C₄₅H₅₀N₃NaO₆S₄, Cat: 152111-91-6) were purchased from Sigma-Aldrich (St.Louis, MO, USA). Cell Counting Kit-8 kit (CCK-8 kit) was from Dojindo Laboratories (Kumamoto, Japan). Annexin V-FITC/PI Apoptosis Detection kit was purchased from Nanjing KeyGen Biotech Co. (Nanjing, China). Hematoxylin-Eosin Staining Kit was from Beyotime Institute of Biotechnology (Shanghai, China). Balb/C nude mice (18-22 g, 5 weeks old, female) were obtained from Comparative Medicine Centre, Yangzhou University (Yangzhou, China). All the experiments were conducted according to the manufacturer's protocols. All reagents were of analytical grade. This study was approved by the Research Ethics Board of Zhongda Hospital affiliated to Southeast University.