Journal:

Article Title: The immunosuppressive drug mycophenolate mofetil impairs the adhesion capacity of gastrointestinal tumour cells

doi: 10.1046/j.1365-2249.2003.02290.x

Figure Lengend Snippet: Fluorescence analysis (relative fluorescence units, RFU) of endothelial cell ICAM-1, VCAM-1, E-selectin and P-selectin surface expression. For maximal stimulation, HUVEC were incubated with IL-1 (100 U/ml) for 12 h to up-regulate ICAM-1, and to induce VCAM-1 and E-selectin. HUVEC were incubated with PGE2 (10−6m) for 10 min to induce P-selectin expression. FL-1H (log) channel histogram analysis; 1 × 104 cells/scan. Control values were set at 100%. Mean ± s.d. of three experiments.

Article Snippet: After blocking, they were incubated overnight with sheep-antihuman P-selectin-antibodies (sheep IgG antibodies; 1 : 500 dilution; R&D Systems).

Techniques: Fluorescence, Expressing, Incubation, Control

Journal:

Article Title: The immunosuppressive drug mycophenolate mofetil impairs the adhesion capacity of gastrointestinal tumour cells

doi: 10.1046/j.1365-2249.2003.02290.x

Figure Lengend Snippet: MMF suppresses IL-1 induced E-selectin mRNA expression. Total RNA was isolated and E-selectin mRNA levels were determined by Northern blot analysis. GAPDH was used to assess equivalent RNA loading C−= unstimulated control cells, C+= IL-1 stimulated control cells. Results of densitometry are shown in the graph below and are expressed as the ratio of E-selectin : GAPDH mRNA, relative to the control (IL-1 activation without MMF), which was assigned a value of 100%. The figure shows one representative experiment from three.

Article Snippet: After blocking, they were incubated overnight with sheep-antihuman P-selectin-antibodies (sheep IgG antibodies; 1 : 500 dilution; R&D Systems).

Techniques: Expressing, Isolation, Northern Blot, Control, Activation Assay

Journal:

Article Title: The immunosuppressive drug mycophenolate mofetil impairs the adhesion capacity of gastrointestinal tumour cells

doi: 10.1046/j.1365-2249.2003.02290.x

Figure Lengend Snippet: Western blot analysis of P-selectin in endothelial cells. HUVEC were stimulated with 10−6m PGE2 and incubated additionally with different concentrations of MMF. Throm = control analysis of platelet P-selectin protein, C-= unstimulated HUVEC without MMF, C+ = PGE2-stimulated HUVEC without MMF. The figure shows one of four separate experiments.

Article Snippet: After blocking, they were incubated overnight with sheep-antihuman P-selectin-antibodies (sheep IgG antibodies; 1 : 500 dilution; R&D Systems).

Techniques: Western Blot, Incubation, Control

Journal:

Article Title: The immunosuppressive drug mycophenolate mofetil impairs the adhesion capacity of gastrointestinal tumour cells

doi: 10.1046/j.1365-2249.2003.02290.x

Figure Lengend Snippet: Adhesion of CD4+ T (a), CD8+ T (b) or WiDr (c) cells to immobilized adhesion receptor globulin chimeras. Purified T cells or WiDr cells were added to culture dishes coated with the extracellular domain of ICAM-1, VCAM-1, E-selectin or P-selectin. The immobilized adhesion proteins were coupled to goat-antihuman IgG. Cells were pretreated with different concentrations of MMF. Untreated cells served as controls (100% value). Adhesion was measured after 30 min incubation and after removal of non-adherent cells, according to the protocol given in Materials and methods. Each point represents the mean ± s.d. of four experiments; s.d. was usually about 20%.

Article Snippet: After blocking, they were incubated overnight with sheep-antihuman P-selectin-antibodies (sheep IgG antibodies; 1 : 500 dilution; R&D Systems).

Techniques: Purification, Incubation

Journal:

Article Title: M-Like Proteins of Streptococcus dysgalactiae

doi:

Figure Lengend Snippet: SDS-PAGE and Western blotting. (a) The affinity-purified GST fusion proteins coded by pGDEMA7 and pGDEMA8 clones derived from S. dysgalactiae strain 8215 were separated on an SDS-PAGE gel (8 to 25% polyacrylamide) (A) and blotted to an NC membrane. Binding of Fg by the recombinant proteins was tested by probing with 125I-Fg and visualized by exposing the strips to x-ray film (B). IgG binding by the recombinant proteins was probed with HRP-labelled human (C) and rabbit (D) IgG while HRP-labelled protein G was used to detect the bound unlabelled horse IgG (E). Lanes: 1, GST; 2, GDEMA7; 3, GDEMA8. (b) Human Fg in reducing sample buffer and Fab and Fc fragments of human IgG in nonreducing sample buffer were separated on a homogeneous SDS–12% PAGE gel (A), transferred to an NC membrane, and probed with 125I-GDEMA8 protein. The bound probe was detected by exposing the membrane to x-ray film (B). Lanes: 1, Fg (from the top: α-, β-, and γ-chains); 2, human IgG Fab fragment; 3, human IgG Fc fragment. Molecular mass markers (kDa) are indicated.

Article Snippet: Rabbit anti-sheep IgG-HRP was the product of Southern Biotechnology Associates, Inc., Birmingham, Ala.

Techniques: SDS Page, Western Blot, Affinity Purification, Clone Assay, Derivative Assay, Binding Assay, Recombinant