Anti rabbit igg fitc

The collected tumors were fixed with 4% paraformaldehyde, embedded in paraffin, and cut into 4‐μm‐thick sections using a microtome. After removing the paraffin wax with xylene, antigens were retrieved with a microwave treatment in 10 mm EDTA buffer (pH 9.0). The tissue sections were blocked with 5% BSA for 1 h at room temperature and were incubated with rabbit polyclonal anti‐human BRF1 antibodies (1 : 200) or Mouse monoclonal ERα antibody (1 : 100) overnight at 4 °C and then incubated with anti‐rabbit IgG FITC or anti‐rabbit IgG CY3 (Invitrogen Life Technologies Corporation, Invitrogen, Carlsbad, CA, USA) as secondary antibodies (1 : 4000). Nuclear staining of cells was performed using 4,6‐diamidino‐2‐phenylindole (DAPI, Beyotime Biotechnology, Shanghai, China). The slides were mounted in antifade reagent (Invitrogen Life Technologies Corporation). The photomicrographs were captured using an Olympus BX63 fluorescence microscopy (Germany).

SB202190 (S7067) was purchased from Sigma-Aldrich (Darmstadt, Germany). MK-2206 was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Dehydroxymethylepoxyquinomicin (DHMEQ) was kindly provided by Prof. Kazuo Umezawa, Aichi Medical University, Japan. Recombinant human (rh) EMMPIN/CD147 Fc chimera (NS0) protein (972-EMN), rhIL-6 (206-IL), rhIL -8 (208-IL), and rhGM-CSF (215-GM) were purchased from R&D system (Minneapolis, MN, USA).
The sources of antibodies were as follows; rabbit anti-NF-κB p65 (C-20, sc-372), polyclonal antibody (pAb), mouse anti-p-p38 (D8, sc-7973) monoclonal antibody (mAb), and mouse anti-p38 (A-12, sc-7972) mAb were from Santa Cruz Biotechnology; rabbit anti-p-Akt (S473, #9271) pAb, rabbit anti-Akt (#9272) pAb, horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (#7074), and HRP-conjugated anti-mouse IgG (#7076) were from Cell Signaling Technology; anti-CD147 (MEM-M6/1) was from Abcam (Cambridge, UK); HRP-conjugated goat anti-mouse IgM was from Southern Biotech (Birmingham, AL, USA, #1020-05); anti-rabbit-IgG-FITC was from Invitrogen (Carlsbad, CA, USA, #656111). All cell culture-related reagents were from Gibco (Thermo Fisher Scientific, Waltham, MA, USA), and chemicals were purchased from Sigma-Aldrich. Otherwise, they will be specified.

Antibodies were purchased as followed: Anti-CD4-V450, anti-CD8α-APCcy7, anti-B220-V450, anti-CD138-Pe-cy7, anti-CD86-Pe-cy5, anti-FAS-PE, anti-GL-7-APC, anti-CD40-APC, anti-MHCII-FITC, anti-IL-4-PE, anti-IL-5-PE, anti-IgM-Pe-cy7, anti-IgG1-APC, anti-IFN-r-Percp5.5, anti-TNF-α-PE, and anti-IL-17-Pe-cy7 were purchased from BD Biosciences (Franklin Lakes, NJ). Anti-PDL-1-biotin (eBioscience, San Diego, CA), anti-XBP-1s (Cell Signaling Technology, Danvers, MA), and anti-Rabbit IgG-FITC (Thermo Fisher Scientific, Waltham, MA) antibodies were purchased from commercial sources. Recombinant mouse IL-4 (PeproTech, Rocky Hill, NJ) and LPS (Sigma-Aldrich, St. Louis, MO) were purchased from the commercial companies. Goat F(ab’)2 Anti-Mouse IgM (1022-01, SouthernBiotech, Birmingham, AL) and anti-mouse CD40 (BE0016-2, BioXCell, Lebanon, NH) were commercially purchased from companies.