Two-cell embryos were collected at day 2 of pregnancy and cultured in single-step medium for another 2-day period. Hatched blastocysts were added to the confluent monolayers of OPN-targeted siRNA or NC-siRNA pre-treated mESC in DMEM/F12 containing 10% cFBS for 48 h. Immunofluorescence was performed on this embryo-mESC co-culture as previously described [29] (link). Cells were incubated with a rabbit E-cadherin antibody (1∶100 dilution, Cell Signaling Technology, Boston, USA), goat polyclonal MMP-9 antibody (1∶100 dilution, Santa Cruz) or rabbit Ig G (1∶100 dilution, Santa Cruz), followed by the FITC conjugated secondary antibody (PIERCE, Rockford, IL, USA). Nuclei were stained with DAPI (Zhongshan Golden Bridge Bio-technology, Beijing, China). The trophoblast spreading area was quantified by measuring the E-cadherin signal area using the Image J software. A total of 10 embryos were calculated in each experiment, and the experiments were repeated three times.
Goat polyclonal mmp 9 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Goat polyclonal MMP-9 antibody is a laboratory reagent used for the detection and analysis of matrix metalloproteinase-9 (MMP-9) in biological samples. MMP-9 is an enzyme involved in the breakdown of extracellular matrix proteins and plays a role in various physiological and pathological processes.
Automatically generated - may contain errors
2 protocols using goat polyclonal mmp 9 antibody
1
Embryo-mESC Co-culture for Trophoblast Spreading
2
Immunohistochemical Analysis of Tumor Samples
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Paraffin-embedded sections were obtained from the tumors treated with BP/LPPC by i.v., i.t. or i.c. injection and were processed for immunohistochemical staining. Briefly, the slides were treated with 3% hydrogen peroxide in 1× PBS for 10 min to block endogenous peroxidase activity after being dewaxed and rehydrated. Next, the sections were washed three times with PBS-T (1×PBS containing 0.1% Tween 20) for 5 min each time, and nonspecific antibody binding was blocked by 10% fetal bovine serum in PBS for 10 min at room temperature. The sections were incubated with a rabbit polyclonal PCNA, caspase 3, VEGF or VEGFR1 (1: 200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA) antibody; a mouse monoclonal VEGFR2 or MMP2 antibody (1: 200 dilution, Santa Cruz Biotechnology); goat polyclonal MMP9 antibody (1: 200 dilution, Santa Cruz Biotechnology) at 4°C overnight, and the immune complexes were probed using a horseradish peroxidase-conjugated anti-mouse, anti-rabbit or anti-goat IgG secondary antibody (1: 1000 dilution; Santa Cruz Biotechnology) and visualized by diaminobenzidine (DAB). Finally, the sections were counterstained with hematoxylin, mounted, observed under a light microscope at a magnification of 400×, and photographed.
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