Incubation:Article Title: Association of hepatocyte-derived growth factor receptor/caudal type homeobox 2 co-expression with mucosal regeneration in active ulcerative colitis
Article Snippet: .. TMA slides and blood smears were incubated with rabbit polyclonal anti-Met culture supernatant antibody (Santa Cruz Biotechnology Inc.; Clone: C-12; 1:100 dilution in PBS), monoclonal mouse anti-human CDX2 antibodies (Clone: AMT28; BioSystems; 1:100 dilution in PBS; Clone: ZC007; Invitrogen; 5 μg/mL working concentration, diluted in PBS) and monoclonal anti-human CD133/1-biotin antibody (Clone: AC133; Miltenyi; 1:100 dilution in PBS) at 37 °C for 60 min. After rinsing 3 times with PBS, samples incubated with anti-Met antibody were finally treated with an antirabbit EnVision polymerHRP conjugate kit (K4003, DAKO) for 40 min. .. Secondary immunodetection in the cases of samples incubated with anti-CDX2 and anti-CD133 antibodies was performed with EnVision System Labelled Polymer-HRP K4001 (Anti-Mouse 1/1; DAKO), as described in the manual.
Article Title: Association of hepatocyte-derived growth factor receptor/caudal type homeobox 2 co-expression with mucosal regeneration in active ulcerative colitis
Article Snippet: Fot the detection of co-expression of the assayed proteins HGFR/CDX2, HGFR/CD133, CD133/CDX2 and Musashi-1/CDX2 double immunofluorescent labelings were performed. .. HGFR/CDX2, HGFR/CD133, Musashi-1/CDX2 labelings: TMA slides and peripheral blood smears were covered with rabbit polyclonal anti-Met culture supernatant antibody (Santa Cruz Biotechnology Inc.; Clone: C-12; 1:100 dilution in PBS) or anti-Musashi-1 monoclonal antibody (EP1302, Abcam, 1:100 dilution in PBS) at 37 °C for 60 min. After rinsing them thrice in PBS, diluted goat polyclonal anti-Rabbit IgG Antibody, biotin-SP conjugate (Millipore Merck; Clone: AP132B; 1:500 dilution in PBS) was added to each sample, and they were incubated at 37 °C for 30 min. After washing them thrice in PBS, samples were covered with diluted Texas Red Streptavidin (1 μL streptavidin in 100 μL of PBS; Jackson Immuno Research Laboratories Inc.) and were also incubated at 37 °C for 30 min. Then samples were incubated with monoclonal mouse anti-human CDX2 antibody (Clone: AMT28; BioSystems; 1:100 dilution in PBS) or monoclonal anti-human CD133/1-biotin antibody (Clone: AC133; Miltenyi; 1:100 dilution in PBS) at 37 °C for 60 min. After rinsing them thrice in PBS, samples were covered with 100x diluted FITC-labeled anti-mouse IgG antibody (Sigma-Aldrich), and were incubated at 37 °C for 30 min. After PBS rinsing, samples were covered with antifading VectaShield (Vector Laboratories Inc.) and coverslips. .. CD133/CDX2 labeling: TMA slides and peripheral blood smears were covered with monoclonal anti-human CD133/1-biotin antibody (Clone: AC133; Miltenyi; 1:100 dilution in PBS) at 37 °C for 60 min. After rinsing them thrice in PBS, diluted Biotin-SP-Conjugated Affinipure Goat anti-mouse IgG (1 μL biotin in 100 μL of PBS; Jackson Immuno Research Laboratories Inc.) was added to each sample, and they were incubated at 37 °C for 30 min. After rinsing them thrice in PBS, samples were covered with diluted Texas Red Streptavidin (1 μL streptavidin in 100 μL of PBS; Jackson Immuno Research Laboratories Inc.), and were incubated at 37 °C for 30 min. After washing them thrice in PBS, samples were incubated with monoclonal mouse anti-human CDX2 antibody (Clone: AMT28; BioSystems; 1:100 dilution in PBS) at 37 °C for 60 min. After rinsing them thrice in PBS, samples were covered with 100x diluted FITC-labeled anti-mouse IgG antibody (Sigma-Aldrich) and were incubated at 37 °C for 30 min. After PBS rinsing, samples were covered with antifading VectaShield (Vector Laboratories Inc.) and coverslips.
Article Title: Survival of HT29 cancer cells is influenced by hepatocyte growth factor receptor inhibition through modulation of self-DNA-triggered TLR9-dependent autophagy response
Article Snippet: .. To detect HGFR, TLR9, and autophagy-associated ATG16L2, Beclin1, and LC3 protein expression, HT29 cell smears were incubated with rabbit polyclonal anti-Met culture supernatant antibody (1:100, Clone: C-12, Santa Cruz Biotechnology Inc.), mouse anti-human monoclonal anti-TLR9 antibody (20 μg/mL; LS-B2341, clone: 26C593.2; LifeSpan BioSciences, USA), and anti-ATG16L1-, anti-BECN1-, and anti-MAP1LC3B antibodies (1:200, Antibody Verify, LA, USA) at 37°C for 1 hour. .. After three rounds of PBS rinsing, cell smears were treated for 40 minutes with an anti-rabbit EnVision polymerHRP conjugate kit (K4003, DAKO).
Concentration Assay:Article Title: Association of hepatocyte-derived growth factor receptor/caudal type homeobox 2 co-expression with mucosal regeneration in active ulcerative colitis
Article Snippet: .. TMA slides and blood smears were incubated with rabbit polyclonal anti-Met culture supernatant antibody (Santa Cruz Biotechnology Inc.; Clone: C-12; 1:100 dilution in PBS), monoclonal mouse anti-human CDX2 antibodies (Clone: AMT28; BioSystems; 1:100 dilution in PBS; Clone: ZC007; Invitrogen; 5 μg/mL working concentration, diluted in PBS) and monoclonal anti-human CD133/1-biotin antibody (Clone: AC133; Miltenyi; 1:100 dilution in PBS) at 37 °C for 60 min. After rinsing 3 times with PBS, samples incubated with anti-Met antibody were finally treated with an antirabbit EnVision polymerHRP conjugate kit (K4003, DAKO) for 40 min. .. Secondary immunodetection in the cases of samples incubated with anti-CDX2 and anti-CD133 antibodies was performed with EnVision System Labelled Polymer-HRP K4001 (Anti-Mouse 1/1; DAKO), as described in the manual.
Expressing:Article Title: Survival of HT29 cancer cells is influenced by hepatocyte growth factor receptor inhibition through modulation of self-DNA-triggered TLR9-dependent autophagy response
Article Snippet: .. To detect HGFR, TLR9, and autophagy-associated ATG16L2, Beclin1, and LC3 protein expression, HT29 cell smears were incubated with rabbit polyclonal anti-Met culture supernatant antibody (1:100, Clone: C-12, Santa Cruz Biotechnology Inc.), mouse anti-human monoclonal anti-TLR9 antibody (20 μg/mL; LS-B2341, clone: 26C593.2; LifeSpan BioSciences, USA), and anti-ATG16L1-, anti-BECN1-, and anti-MAP1LC3B antibodies (1:200, Antibody Verify, LA, USA) at 37°C for 1 hour. .. After three rounds of PBS rinsing, cell smears were treated for 40 minutes with an anti-rabbit EnVision polymerHRP conjugate kit (K4003, DAKO).
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