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anti human cd59 mabs  (Bio-Rad)
93
Bio-Rad anti human cd59 mabs
FIGURE 2. miR-200 (b and c) regulates cell sensitivity to complement, C5b-9 deposition, and CD46 and CD55 expression. (A) K562 cells were transfected with miR-200b/c or control plasmid. After 3 or 24 h, cells were treated with Ab and NHS. Cell death (%) was measured by trypan blue inclusion. (B) K562 cells were transfected with miRNA inhibitors specific for miR-200b and miR-200c, or with nonspecific oligonucleotide as negative control. After 24 h, cells were treated with Ab and complement, as above. Cell death (%) was measured by propidium iodide inclusion. (C) K562 cells transfected with miR-200b/c or a control plasmid for 24 h were treated with a sublytic dose of Ab and then with NHS for 10 min at 37˚C. Then they were labeled with goat anti-C3 Ab and FITC-conjugated secondary Ab and analyzed by flow cytometry. Mean fluorescence intensity (MFI) values of cell-bound C3b, representative of three independent experiments, are shown. (D) K562 cells transfected with miR-200b/c or a control plasmid for 24 h were treated with sublytic complement, as in (C). Then they were labeled with mouse anti– C5b-9 aE-11 Ab and FITC-conjugated secondary Ab and analyzed by flow cytometry. MFI values of cell-bound C5b-9, representative of three independent experiments, are shown. (E) K562 cells transfected for 24 h with inhibitors of miR-200b and miR-200c, or with nonspecific oligonucleotide, were incubated with sublytic Ab and NHS for 10 min at 37˚C. The cells were labeled with the aE-11 Ab, as above. MFI values of cell-bound C5b-9, representative of three independent experiments, are shown. (F) K562 cells were transfected with miR-200b/c or control plasmid for 24 h and were labeled with mouse anti-CD46, anti-CD55, or anti- <t>CD59</t> Ab and then with FITC-conjugated secondary Ab. Cells were then analyzed by flow cytometry, and MFI values were determined. Results, representative of three independent experiments, present each regulator’s level relative to its expression in control cells (set as 100%). *p , 0.05, **p , 0.01 relative to control.
Anti Human Cd59 Mabs, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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FIGURE 2. miR-200 (b and c) regulates cell sensitivity to complement, C5b-9 deposition, and CD46 and CD55 expression. (A) K562 cells were transfected with miR-200b/c or control plasmid. After 3 or 24 h, cells were treated with Ab and NHS. Cell death (%) was measured by trypan blue inclusion. (B) K562 cells were transfected with miRNA inhibitors specific for miR-200b and miR-200c, or with nonspecific oligonucleotide as negative control. After 24 h, cells were treated with Ab and complement, as above. Cell death (%) was measured by propidium iodide inclusion. (C) K562 cells transfected with miR-200b/c or a control plasmid for 24 h were treated with a sublytic dose of Ab and then with NHS for 10 min at 37˚C. Then they were labeled with goat anti-C3 Ab and FITC-conjugated secondary Ab and analyzed by flow cytometry. Mean fluorescence intensity (MFI) values of cell-bound C3b, representative of three independent experiments, are shown. (D) K562 cells transfected with miR-200b/c or a control plasmid for 24 h were treated with sublytic complement, as in (C). Then they were labeled with mouse anti– C5b-9 aE-11 Ab and FITC-conjugated secondary Ab and analyzed by flow cytometry. MFI values of cell-bound C5b-9, representative of three independent experiments, are shown. (E) K562 cells transfected for 24 h with inhibitors of miR-200b and miR-200c, or with nonspecific oligonucleotide, were incubated with sublytic Ab and NHS for 10 min at 37˚C. The cells were labeled with the aE-11 Ab, as above. MFI values of cell-bound C5b-9, representative of three independent experiments, are shown. (F) K562 cells were transfected with miR-200b/c or control plasmid for 24 h and were labeled with mouse anti-CD46, anti-CD55, or anti- CD59 Ab and then with FITC-conjugated secondary Ab. Cells were then analyzed by flow cytometry, and MFI values were determined. Results, representative of three independent experiments, present each regulator’s level relative to its expression in control cells (set as 100%). *p , 0.05, **p , 0.01 relative to control.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Regulation of Complement-Dependent Cytotoxicity by MicroRNAs miR-200b, miR-200c, and miR-217.

doi: 10.4049/jimmunol.1502701

Figure Lengend Snippet: FIGURE 2. miR-200 (b and c) regulates cell sensitivity to complement, C5b-9 deposition, and CD46 and CD55 expression. (A) K562 cells were transfected with miR-200b/c or control plasmid. After 3 or 24 h, cells were treated with Ab and NHS. Cell death (%) was measured by trypan blue inclusion. (B) K562 cells were transfected with miRNA inhibitors specific for miR-200b and miR-200c, or with nonspecific oligonucleotide as negative control. After 24 h, cells were treated with Ab and complement, as above. Cell death (%) was measured by propidium iodide inclusion. (C) K562 cells transfected with miR-200b/c or a control plasmid for 24 h were treated with a sublytic dose of Ab and then with NHS for 10 min at 37˚C. Then they were labeled with goat anti-C3 Ab and FITC-conjugated secondary Ab and analyzed by flow cytometry. Mean fluorescence intensity (MFI) values of cell-bound C3b, representative of three independent experiments, are shown. (D) K562 cells transfected with miR-200b/c or a control plasmid for 24 h were treated with sublytic complement, as in (C). Then they were labeled with mouse anti– C5b-9 aE-11 Ab and FITC-conjugated secondary Ab and analyzed by flow cytometry. MFI values of cell-bound C5b-9, representative of three independent experiments, are shown. (E) K562 cells transfected for 24 h with inhibitors of miR-200b and miR-200c, or with nonspecific oligonucleotide, were incubated with sublytic Ab and NHS for 10 min at 37˚C. The cells were labeled with the aE-11 Ab, as above. MFI values of cell-bound C5b-9, representative of three independent experiments, are shown. (F) K562 cells were transfected with miR-200b/c or control plasmid for 24 h and were labeled with mouse anti-CD46, anti-CD55, or anti- CD59 Ab and then with FITC-conjugated secondary Ab. Cells were then analyzed by flow cytometry, and MFI values were determined. Results, representative of three independent experiments, present each regulator’s level relative to its expression in control cells (set as 100%). *p , 0.05, **p , 0.01 relative to control.

Article Snippet: Mouse anti-human CD46, anti-human CD55, and anti-human CD59 mAbs were purchased from AbD Serotec (Oxford, U.K.).

Techniques: Expressing, Transfection, Control, Plasmid Preparation, Negative Control, Labeling, Cytometry, Incubation

FIGURE 3. miR-217 regulates cell sensitivity to complement, C3 and C5b-9 deposition, and CD46 expression. (A) K562 cells were transfected with miR-217 or control plasmid. After 3 or 24 h, cells were treated with Ab and NHS (60 min). Cell death (%) was measured by trypan blue inclusion. (B) K562 cells were transfected with miRNA inhibitors specific for miR-217 or with nonspecific oligonucleotide as negative control. After 24 h, cells were treated with Ab and complement, as above. Cell death (%) was measured by propidium iodide inclusion. (C and D) K562 cells transfected for 24 h with miR-217 or control plasmid were incubated with a sublytic dose of Ab and NHS for 10 min at 37˚C. Cells were labeled with goat anti-C3b Ab and FITC-conjugated secondary Ab (C) or mouse anti–C5b-9 aE-11 Ab and FITC-conjugated secondary Ab (D) and analyzed by flow cytometry. Mean fluorescence intensity (MFI) values of cell-bound C3b or C5b-9, representative of three independent experiments, are shown. (E) K562 cells transfected for 24 h with miR-217 inhibitor or with nonspecific oligonucleotide (C) were incubated with a sublytic dose of Ab and NHS for 10 min at 37˚C. The cells were labeled with aE-11 Ab and FITC-conjugated secondary Ab, and MFI values of cell-bound C5b-9 were determined. Results are representative of three independent experiments. (F) K562 cells were transfected with miR-217 or control plasmid for 24 h and were then labeled with mouse anti-CD46, anti-CD55, or anti-CD59 Ab and FITC-conjugated secondary Ab. Cells were then analyzed by flow cytometry, and MFI values were determined. Results, representative of three independent experiments, present each regulator’s level relative to its expression in control cells (set as 100%). *p , 0.05, **p , 0.01 relative to control.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Regulation of Complement-Dependent Cytotoxicity by MicroRNAs miR-200b, miR-200c, and miR-217.

doi: 10.4049/jimmunol.1502701

Figure Lengend Snippet: FIGURE 3. miR-217 regulates cell sensitivity to complement, C3 and C5b-9 deposition, and CD46 expression. (A) K562 cells were transfected with miR-217 or control plasmid. After 3 or 24 h, cells were treated with Ab and NHS (60 min). Cell death (%) was measured by trypan blue inclusion. (B) K562 cells were transfected with miRNA inhibitors specific for miR-217 or with nonspecific oligonucleotide as negative control. After 24 h, cells were treated with Ab and complement, as above. Cell death (%) was measured by propidium iodide inclusion. (C and D) K562 cells transfected for 24 h with miR-217 or control plasmid were incubated with a sublytic dose of Ab and NHS for 10 min at 37˚C. Cells were labeled with goat anti-C3b Ab and FITC-conjugated secondary Ab (C) or mouse anti–C5b-9 aE-11 Ab and FITC-conjugated secondary Ab (D) and analyzed by flow cytometry. Mean fluorescence intensity (MFI) values of cell-bound C3b or C5b-9, representative of three independent experiments, are shown. (E) K562 cells transfected for 24 h with miR-217 inhibitor or with nonspecific oligonucleotide (C) were incubated with a sublytic dose of Ab and NHS for 10 min at 37˚C. The cells were labeled with aE-11 Ab and FITC-conjugated secondary Ab, and MFI values of cell-bound C5b-9 were determined. Results are representative of three independent experiments. (F) K562 cells were transfected with miR-217 or control plasmid for 24 h and were then labeled with mouse anti-CD46, anti-CD55, or anti-CD59 Ab and FITC-conjugated secondary Ab. Cells were then analyzed by flow cytometry, and MFI values were determined. Results, representative of three independent experiments, present each regulator’s level relative to its expression in control cells (set as 100%). *p , 0.05, **p , 0.01 relative to control.

Article Snippet: Mouse anti-human CD46, anti-human CD55, and anti-human CD59 mAbs were purchased from AbD Serotec (Oxford, U.K.).

Techniques: Expressing, Transfection, Control, Plasmid Preparation, Negative Control, Incubation, Labeling, Cytometry