mouse anti human cd55 Search Results


mouse anti human cd55  (Bio-Rad)
93
Bio-Rad mouse anti human cd55
FIGURE 4. CML triggered by anti–EGFR-IgG3 negatively correlates with <t>CD55</t> and CD59 expression levels. (A) Surface expression levels of CD46, CD55, and CD59 on analyzed cell lines were quantified by calibrated flow cytometry. Means 6 SEM of at least three independent experiments are presented. (B) Correlations between CD46, CD55, or CD59 and anti–EGFR-IgG3–mediated CDC were calculated for all four cell lines. CDC results at 2 mg/ml Ab concentration were taken from experiments presented in Fig. 2. (C) A431 cells were seeded into 10-cm plates and grown overnight. On the following day, cells were transfected with 50 nM control siRNA or with single siRNAs specific for CD46, CD55, CD59, or with a combination of all three mCRP-specific siRNAs for 72 h. Efficiency of siRNA-induced knockdown was analyzed by direct flow cytometry using fluorochrome-labeled, mCRP- specific Abs (CD46-Pacific blue, CD55-PE, CD59-FITC), or respective control Abs. (D–G) CDC against control siRNA or mCRP-specific, siRNA- transfected A431 cells was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human serum and <t>anti–EGFR-IgG1</t> (upper panels), anti– EGFR-IgG3 (lower panels), as well as the respective control Abs at increasing concentrations. (H) Concentration-dependent binding of CD55-Ab <t>(BRIC216,</t> mouse IgG1) to A431 cells was analyzed by indirect immunofluorescence. Results from one representative experiment are presented. (I) CDC triggered by anti–EGFR-IgG1, anti–EGFR-IgG3, or respective control Abs (all at 66.67 nM) against A431 cells in the presence of saturating concentrations of CD55-Ab or a control Ab (both at 66.67 nM) was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human (Figure legend continues)
Mouse Anti Human Cd55, 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
https://www.bioz.com/result/mouse anti human cd55/product/Bio-Rad
Average 93 stars, based on 1 article reviews
mouse anti human cd55 - by Bioz Stars, 2026-04
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fluorescein isothiocyanate (fitc)-conjugated mouse anti-human cd55  (4A Biotech)
90
4A Biotech fluorescein isothiocyanate (fitc)-conjugated mouse anti-human cd55
Expression of <t>CD55</t> and CD59 in tumor cells. (A) Cell lines were incubated with fluorescein isothiocyanate-conjugated anti-human CD55 and CD59 monoclonal antibodies, and analyzed by flow cytometry. Error bars depict standard deviations. (B) The protein level of CD55 and CD59 in A549 and Lovo cells were detected by western blotting. β-actin protein levels served as the loading control. CD55, <t>decay</t> <t>accelerating</t> <t>factor;</t> CD59, protectin.
Fluorescein Isothiocyanate (Fitc) Conjugated Mouse Anti Human Cd55, supplied by 4A Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/fluorescein isothiocyanate (fitc)-conjugated mouse anti-human cd55/product/4A Biotech
Average 90 stars, based on 1 article reviews
fluorescein isothiocyanate (fitc)-conjugated mouse anti-human cd55 - by Bioz Stars, 2026-04
90/100 stars
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primary mouse anti-human cd55 antibody (bric110)  (Cymbus Bioscience Ltd)
90
Cymbus Bioscience Ltd primary mouse anti-human cd55 antibody (bric110)
hCD55 and α-Gal expression on porcine cells. (a) Expression of <t>CD55</t> was analyzed by FACs on both transgenic (TgPAE) and nontransgenic (PAE) pig cells. Staining by anti-hCD55 (shaded histogram), no primary antibody (negative control; solid line), or anti-hCD46 (dotted line, similar profile to that of the no-antibody control) followed by FITC-labeled secondary anti-mouse immunoglobulin antibodies (upper panels) is shown. Staining for α-Gal was a one-step incubation; hence, −lectin on the histogram represents cells only, and +lectin represents cells incubated with IB-4 lectin conjugated to FITC (lower panels). (b) α-Gal expression on ST-IOWA wild-type and ST-IOWA Gal-null (−/−) cells. Expression of the α-Gal antigen was analyzed by FACs, using the IB-4 lectin. A rightward shift of the histogram in the presence of lectin (+lectin) compared to the histogram generated in the absence of lectin (−lectin) indicates expression of the α-Gal antigen. The level of expression was determined using the MFI shift, generated by the Cell Quest FACScan software. The MFI shift was calculated by determining test MFI and subtracting that of negative controls (no primary antibodies for hCD55 and hCD46 expression and no lectin for α-Gal expression), and all the mean shifts are shown in Table ​Table11.
Primary Mouse Anti Human Cd55 Antibody (Bric110), supplied by Cymbus Bioscience Ltd, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/primary mouse anti-human cd55 antibody (bric110)/product/Cymbus Bioscience Ltd
Average 90 stars, based on 1 article reviews
primary mouse anti-human cd55 antibody (bric110) - by Bioz Stars, 2026-04
90/100 stars
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bv480 mouse anti human cd55  (BD Biosciences)
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apc mouse anti human cd55  (BD Biosciences)
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buv496 mouse anti human cd55  (BD Biosciences)
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mouse anti human cd55 rpe  (RayBiotech)
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Mouse Anti Human CD55 RPE 25 TESTS 0 25ml
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bv786 mouse anti human cd55  (BD Biosciences)
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buv737 mouse anti human cd55  (BD Biosciences)
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buv615 mouse anti human cd55  (BD Biosciences)
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mouse monoclonal anti human cd55 antibody  (Angio-Proteomie)
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mouse anti human cd55 antibody daf  (RayBiotech)
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Mouse anti Human CD55 Antibody DAF
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FIGURE 4. CML triggered by anti–EGFR-IgG3 negatively correlates with CD55 and CD59 expression levels. (A) Surface expression levels of CD46, CD55, and CD59 on analyzed cell lines were quantified by calibrated flow cytometry. Means 6 SEM of at least three independent experiments are presented. (B) Correlations between CD46, CD55, or CD59 and anti–EGFR-IgG3–mediated CDC were calculated for all four cell lines. CDC results at 2 mg/ml Ab concentration were taken from experiments presented in Fig. 2. (C) A431 cells were seeded into 10-cm plates and grown overnight. On the following day, cells were transfected with 50 nM control siRNA or with single siRNAs specific for CD46, CD55, CD59, or with a combination of all three mCRP-specific siRNAs for 72 h. Efficiency of siRNA-induced knockdown was analyzed by direct flow cytometry using fluorochrome-labeled, mCRP- specific Abs (CD46-Pacific blue, CD55-PE, CD59-FITC), or respective control Abs. (D–G) CDC against control siRNA or mCRP-specific, siRNA- transfected A431 cells was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human serum and anti–EGFR-IgG1 (upper panels), anti– EGFR-IgG3 (lower panels), as well as the respective control Abs at increasing concentrations. (H) Concentration-dependent binding of CD55-Ab (BRIC216, mouse IgG1) to A431 cells was analyzed by indirect immunofluorescence. Results from one representative experiment are presented. (I) CDC triggered by anti–EGFR-IgG1, anti–EGFR-IgG3, or respective control Abs (all at 66.67 nM) against A431 cells in the presence of saturating concentrations of CD55-Ab or a control Ab (both at 66.67 nM) was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human (Figure legend continues)

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

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 4. CML triggered by anti–EGFR-IgG3 negatively correlates with CD55 and CD59 expression levels. (A) Surface expression levels of CD46, CD55, and CD59 on analyzed cell lines were quantified by calibrated flow cytometry. Means 6 SEM of at least three independent experiments are presented. (B) Correlations between CD46, CD55, or CD59 and anti–EGFR-IgG3–mediated CDC were calculated for all four cell lines. CDC results at 2 mg/ml Ab concentration were taken from experiments presented in Fig. 2. (C) A431 cells were seeded into 10-cm plates and grown overnight. On the following day, cells were transfected with 50 nM control siRNA or with single siRNAs specific for CD46, CD55, CD59, or with a combination of all three mCRP-specific siRNAs for 72 h. Efficiency of siRNA-induced knockdown was analyzed by direct flow cytometry using fluorochrome-labeled, mCRP- specific Abs (CD46-Pacific blue, CD55-PE, CD59-FITC), or respective control Abs. (D–G) CDC against control siRNA or mCRP-specific, siRNA- transfected A431 cells was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human serum and anti–EGFR-IgG1 (upper panels), anti– EGFR-IgG3 (lower panels), as well as the respective control Abs at increasing concentrations. (H) Concentration-dependent binding of CD55-Ab (BRIC216, mouse IgG1) to A431 cells was analyzed by indirect immunofluorescence. Results from one representative experiment are presented. (I) CDC triggered by anti–EGFR-IgG1, anti–EGFR-IgG3, or respective control Abs (all at 66.67 nM) against A431 cells in the presence of saturating concentrations of CD55-Ab or a control Ab (both at 66.67 nM) was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human (Figure legend continues)

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Expressing, Cytometry, Concentration Assay, Transfection, Control, Knockdown, Labeling, Binding Assay

FIGURE 5. CD55 dampens anti–EGFR-IgG3–triggered CML and promotes C1q-dependent induction of AP amplification. BHK-EGFR+ #5 cells were transiently transfected with a control vector or a CD55 vector for 48 h. (A) Cell-surface expression of CD55 was analyzed by direct flow cytometry using PE-conjugated CD55-specific or control Abs. (B–D) The influence of CD55 overexpression on anti–EGFR-IgG3–mediated CDC was investigated by [51Cr] release assays either (B) in an Ab concentration–response curve, (C) in a time-dependent manner, or (D) in serum titration experiments. (E–H) The influence of the alternative complement pathway inhibitor CRIg (E and G), the presence of C1q in serum (F; at 66.67 nM Ab concentration; mean 6 SEM of triplicates), as well as of factor B (H; 13.33 nM Ab concentration, 12.5% v/v factor B–depleted serum, 200 mg/ml factor B), on anti–EGFR-IgG3–mediated CDC was analyzed using either (E and F) control vector–transfected or CD55 vector–transfected BHK-EGFR+ #5 cells or (G and H) DiFi cells (66.67 nM Ab concentration). (I) Deposition of factor Bb on control vector– or CD55 vector–transfected BHK-EGFR+ #5 cells was analyzed by flow cytometry. Relative deposition levels were calculated by equating RFI measured in the absence of Ab with 100%. Results are presented as mean 6 SEM of at least three independent experiments with different blood donors. *p # 0.05 anti–EGFR-IgG3 versus respective control Ab; (B–D, I) #p # 0.05 control vector versus CD55 vector; (E and G) #p # 0.05 without CRIg-Fc versus CRIg-Fc; (H) #p # 0.05 w/o factor B versus with factor B.

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

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 5. CD55 dampens anti–EGFR-IgG3–triggered CML and promotes C1q-dependent induction of AP amplification. BHK-EGFR+ #5 cells were transiently transfected with a control vector or a CD55 vector for 48 h. (A) Cell-surface expression of CD55 was analyzed by direct flow cytometry using PE-conjugated CD55-specific or control Abs. (B–D) The influence of CD55 overexpression on anti–EGFR-IgG3–mediated CDC was investigated by [51Cr] release assays either (B) in an Ab concentration–response curve, (C) in a time-dependent manner, or (D) in serum titration experiments. (E–H) The influence of the alternative complement pathway inhibitor CRIg (E and G), the presence of C1q in serum (F; at 66.67 nM Ab concentration; mean 6 SEM of triplicates), as well as of factor B (H; 13.33 nM Ab concentration, 12.5% v/v factor B–depleted serum, 200 mg/ml factor B), on anti–EGFR-IgG3–mediated CDC was analyzed using either (E and F) control vector–transfected or CD55 vector–transfected BHK-EGFR+ #5 cells or (G and H) DiFi cells (66.67 nM Ab concentration). (I) Deposition of factor Bb on control vector– or CD55 vector–transfected BHK-EGFR+ #5 cells was analyzed by flow cytometry. Relative deposition levels were calculated by equating RFI measured in the absence of Ab with 100%. Results are presented as mean 6 SEM of at least three independent experiments with different blood donors. *p # 0.05 anti–EGFR-IgG3 versus respective control Ab; (B–D, I) #p # 0.05 control vector versus CD55 vector; (E and G) #p # 0.05 without CRIg-Fc versus CRIg-Fc; (H) #p # 0.05 w/o factor B versus with factor B.

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Transfection, Control, Plasmid Preparation, Expressing, Cytometry, Over Expression, Concentration Assay, Titration

FIGURE 6. Overview of complement activation by human anti–EGFR-IgG3 in the context of CD55 expression. On CD55-deficient target cells (left panel), anti–EGFR-IgG3 mediates strong C3b but low C4b deposition and induces assembly of classical and alternative C3 convertases, predominantly resulting in the induction of fast and efficient CDC via the classical pathway of complement activation. In contrast, on CD55-expressing target cells (right panel), CD55 mainly accelerates the decay of low amounts of classical C3 convertases, leading to amplification of the AP and finally to slow and inefficient CDC induction.

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

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 6. Overview of complement activation by human anti–EGFR-IgG3 in the context of CD55 expression. On CD55-deficient target cells (left panel), anti–EGFR-IgG3 mediates strong C3b but low C4b deposition and induces assembly of classical and alternative C3 convertases, predominantly resulting in the induction of fast and efficient CDC via the classical pathway of complement activation. In contrast, on CD55-expressing target cells (right panel), CD55 mainly accelerates the decay of low amounts of classical C3 convertases, leading to amplification of the AP and finally to slow and inefficient CDC induction.

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Activation Assay, Expressing

Expression of CD55 and CD59 in tumor cells. (A) Cell lines were incubated with fluorescein isothiocyanate-conjugated anti-human CD55 and CD59 monoclonal antibodies, and analyzed by flow cytometry. Error bars depict standard deviations. (B) The protein level of CD55 and CD59 in A549 and Lovo cells were detected by western blotting. β-actin protein levels served as the loading control. CD55, decay accelerating factor; CD59, protectin.

Journal: Oncology Letters

Article Title: Effect of membrane-bound complement regulatory proteins on tumor cell sensitivity to complement-dependent cytolysis triggered by heterologous expression of the α-gal xenoantigen

doi: 10.3892/ol.2018.8478

Figure Lengend Snippet: Expression of CD55 and CD59 in tumor cells. (A) Cell lines were incubated with fluorescein isothiocyanate-conjugated anti-human CD55 and CD59 monoclonal antibodies, and analyzed by flow cytometry. Error bars depict standard deviations. (B) The protein level of CD55 and CD59 in A549 and Lovo cells were detected by western blotting. β-actin protein levels served as the loading control. CD55, decay accelerating factor; CD59, protectin.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD55 (cat. no. FHF055) and CD59 (cat. no. FHF059) mAbs were purchased from Beijing 4A Biotech Co., Ltd. (Beijing, China).

Techniques: Expressing, Incubation, Bioprocessing, Flow Cytometry, Western Blot, Control

Establishing stable transfected α-gal-expressing cell lines. (A) α-1,3GT mRNA expression in A549, A549-V, A549-GT, Lovo, Lovo-V and Lovo-GT cells were detected by reverse transcription-polymerase chain reaction. The amplified product of α-1,3GT was detected by agarose gel electrophoresis in lane 2, 4 and 6 of the two gels. GAPDH was used as loading control in lane 1, 3 and 5 of the two gels. (B) Expression of α-gal epitope in each group of cells were detected by direct immunofluorescence (magnification, ×200). FITC-conjugated BS-IB4 lectin staining was performed to probe α-gal epitope. (B-a) A549-GT, (B-b) A549, (B-c) A549-V, (B-d) Lovo-GT, (B-e) Lovo, (B-f) Lovo-V and (B-g) positive control PIEC cells. (C) Expression of α-gal epitope in each group of cells were stained with FITC-BS-IB4 lectin, then analyzed by flow cytometry. Error bars depict standard deviations. FITC, fluorescein isothiocyanate; α-gal, Galα1-3Galβ1-4GlcNAc-R; α-1,3GT, α1,3-galactosyltransferase; PIEC, pig iliac arterial endothelial cells; A549-GT, α-gal expressing A549; A549-V, control.

Journal: Oncology Letters

Article Title: Effect of membrane-bound complement regulatory proteins on tumor cell sensitivity to complement-dependent cytolysis triggered by heterologous expression of the α-gal xenoantigen

doi: 10.3892/ol.2018.8478

Figure Lengend Snippet: Establishing stable transfected α-gal-expressing cell lines. (A) α-1,3GT mRNA expression in A549, A549-V, A549-GT, Lovo, Lovo-V and Lovo-GT cells were detected by reverse transcription-polymerase chain reaction. The amplified product of α-1,3GT was detected by agarose gel electrophoresis in lane 2, 4 and 6 of the two gels. GAPDH was used as loading control in lane 1, 3 and 5 of the two gels. (B) Expression of α-gal epitope in each group of cells were detected by direct immunofluorescence (magnification, ×200). FITC-conjugated BS-IB4 lectin staining was performed to probe α-gal epitope. (B-a) A549-GT, (B-b) A549, (B-c) A549-V, (B-d) Lovo-GT, (B-e) Lovo, (B-f) Lovo-V and (B-g) positive control PIEC cells. (C) Expression of α-gal epitope in each group of cells were stained with FITC-BS-IB4 lectin, then analyzed by flow cytometry. Error bars depict standard deviations. FITC, fluorescein isothiocyanate; α-gal, Galα1-3Galβ1-4GlcNAc-R; α-1,3GT, α1,3-galactosyltransferase; PIEC, pig iliac arterial endothelial cells; A549-GT, α-gal expressing A549; A549-V, control.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD55 (cat. no. FHF055) and CD59 (cat. no. FHF059) mAbs were purchased from Beijing 4A Biotech Co., Ltd. (Beijing, China).

Techniques: Transfection, Expressing, Reverse Transcription, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Control, Immunofluorescence, Staining, Positive Control, Flow Cytometry

Expression of CD55 and CD59 on α-gal-expressing cells influences their sensitivity to CDC. (A) A549, A549-V, A549-GT, Lovo, Lovo-V, Lovo-GT and positive control PIEC cells were incubated with various dilutions of NHS (0, 15, 30, 50%) and survival rates were analyzed by trypan blue staining. Error bars showed standard deviations (*P<0.05 vs. the control). (B) A549, A549-V, A549-GT Cells were pre-treated with various concentrations of PI-PLC (0.001, 0.01, 0.05, 0.1, 0.2, or 0.5 U/ml), incubated with 50% NHS, and survival rates were analyzed by trypan blue staining. Error bars showed standard deviations. *P<0.05, vs. the control. CD55, decay accelerating factor; CD59, protectin; NHS, normal human serum; PI-PLC, phosphatidylinositol-specific phospholipase C; A549-GT, α-gal expressing A549; A549-V, control.

Journal: Oncology Letters

Article Title: Effect of membrane-bound complement regulatory proteins on tumor cell sensitivity to complement-dependent cytolysis triggered by heterologous expression of the α-gal xenoantigen

doi: 10.3892/ol.2018.8478

Figure Lengend Snippet: Expression of CD55 and CD59 on α-gal-expressing cells influences their sensitivity to CDC. (A) A549, A549-V, A549-GT, Lovo, Lovo-V, Lovo-GT and positive control PIEC cells were incubated with various dilutions of NHS (0, 15, 30, 50%) and survival rates were analyzed by trypan blue staining. Error bars showed standard deviations (*P<0.05 vs. the control). (B) A549, A549-V, A549-GT Cells were pre-treated with various concentrations of PI-PLC (0.001, 0.01, 0.05, 0.1, 0.2, or 0.5 U/ml), incubated with 50% NHS, and survival rates were analyzed by trypan blue staining. Error bars showed standard deviations. *P<0.05, vs. the control. CD55, decay accelerating factor; CD59, protectin; NHS, normal human serum; PI-PLC, phosphatidylinositol-specific phospholipase C; A549-GT, α-gal expressing A549; A549-V, control.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD55 (cat. no. FHF055) and CD59 (cat. no. FHF059) mAbs were purchased from Beijing 4A Biotech Co., Ltd. (Beijing, China).

Techniques: Expressing, Positive Control, Incubation, Staining, Control

Effects of PI-PLC treatment on CD55 and CD59 protein level in A549-GT cells. (A) Following 0.1 U/ml PI-PLC treatment, CD55 and CD59 were tested by western blot in A549, A549-V, A549-GT, Lovo, Lovo-V and Lovo-GT cells, compared with that prior to PI-PLC treatment. (B) After 0.1 U/ml PI-PLC treatment, A549-GT cells was incubated with fluorescein isothiocyanate-conjugated anti-human monoclonal antibodies. CD55 and CD59 were analyzed by flow cytometry, compared with that prior to PI-PLC treatment. Error bars showed standard deviations. *P<0.05 vs. the control. CD55, decay accelerating factor; CD59, protectin; PI-PLC, phosphatidylinositol-specific phospholipase C; A549-GT, α-gal expressing A549; A549-V, control.

Journal: Oncology Letters

Article Title: Effect of membrane-bound complement regulatory proteins on tumor cell sensitivity to complement-dependent cytolysis triggered by heterologous expression of the α-gal xenoantigen

doi: 10.3892/ol.2018.8478

Figure Lengend Snippet: Effects of PI-PLC treatment on CD55 and CD59 protein level in A549-GT cells. (A) Following 0.1 U/ml PI-PLC treatment, CD55 and CD59 were tested by western blot in A549, A549-V, A549-GT, Lovo, Lovo-V and Lovo-GT cells, compared with that prior to PI-PLC treatment. (B) After 0.1 U/ml PI-PLC treatment, A549-GT cells was incubated with fluorescein isothiocyanate-conjugated anti-human monoclonal antibodies. CD55 and CD59 were analyzed by flow cytometry, compared with that prior to PI-PLC treatment. Error bars showed standard deviations. *P<0.05 vs. the control. CD55, decay accelerating factor; CD59, protectin; PI-PLC, phosphatidylinositol-specific phospholipase C; A549-GT, α-gal expressing A549; A549-V, control.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD55 (cat. no. FHF055) and CD59 (cat. no. FHF059) mAbs were purchased from Beijing 4A Biotech Co., Ltd. (Beijing, China).

Techniques: Western Blot, Incubation, Bioprocessing, Flow Cytometry, Control, Expressing

Effect of anti-CD55 and anti-CD59 on CDC in α-gal-expressing cells. The A549, A549-V and A549-GT cells were pre-incubated with each antibodies (anti-CD55, anti-CD59 and anti-CD55 with anti-CD59) (10 µg/ml), then 50% normal human serum was added and the survival rates were calculated. Error bars showed standard deviations. *P<0.05 vs. the control. CD55, decay accelerating factor; CD59, protectin; α-gal, Galα1-3Galβ1-4GlcNAc-R; A549-GT, α-gal expressing A549; A549-V, control.

Journal: Oncology Letters

Article Title: Effect of membrane-bound complement regulatory proteins on tumor cell sensitivity to complement-dependent cytolysis triggered by heterologous expression of the α-gal xenoantigen

doi: 10.3892/ol.2018.8478

Figure Lengend Snippet: Effect of anti-CD55 and anti-CD59 on CDC in α-gal-expressing cells. The A549, A549-V and A549-GT cells were pre-incubated with each antibodies (anti-CD55, anti-CD59 and anti-CD55 with anti-CD59) (10 µg/ml), then 50% normal human serum was added and the survival rates were calculated. Error bars showed standard deviations. *P<0.05 vs. the control. CD55, decay accelerating factor; CD59, protectin; α-gal, Galα1-3Galβ1-4GlcNAc-R; A549-GT, α-gal expressing A549; A549-V, control.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD55 (cat. no. FHF055) and CD59 (cat. no. FHF059) mAbs were purchased from Beijing 4A Biotech Co., Ltd. (Beijing, China).

Techniques: Expressing, Incubation, Control

hCD55 and α-Gal expression on porcine cells. (a) Expression of CD55 was analyzed by FACs on both transgenic (TgPAE) and nontransgenic (PAE) pig cells. Staining by anti-hCD55 (shaded histogram), no primary antibody (negative control; solid line), or anti-hCD46 (dotted line, similar profile to that of the no-antibody control) followed by FITC-labeled secondary anti-mouse immunoglobulin antibodies (upper panels) is shown. Staining for α-Gal was a one-step incubation; hence, −lectin on the histogram represents cells only, and +lectin represents cells incubated with IB-4 lectin conjugated to FITC (lower panels). (b) α-Gal expression on ST-IOWA wild-type and ST-IOWA Gal-null (−/−) cells. Expression of the α-Gal antigen was analyzed by FACs, using the IB-4 lectin. A rightward shift of the histogram in the presence of lectin (+lectin) compared to the histogram generated in the absence of lectin (−lectin) indicates expression of the α-Gal antigen. The level of expression was determined using the MFI shift, generated by the Cell Quest FACScan software. The MFI shift was calculated by determining test MFI and subtracting that of negative controls (no primary antibodies for hCD55 and hCD46 expression and no lectin for α-Gal expression), and all the mean shifts are shown in Table ​Table11.

Journal:

Article Title: Reduced Sensitivity to Human Serum Inactivation of Enveloped Viruses Produced by Pig Cells Transgenic for Human CD55 or Deficient for the Galactosyl-?(1-3) Galactosyl Epitope

doi: 10.1128/JVI.78.11.5812-5819.2004

Figure Lengend Snippet: hCD55 and α-Gal expression on porcine cells. (a) Expression of CD55 was analyzed by FACs on both transgenic (TgPAE) and nontransgenic (PAE) pig cells. Staining by anti-hCD55 (shaded histogram), no primary antibody (negative control; solid line), or anti-hCD46 (dotted line, similar profile to that of the no-antibody control) followed by FITC-labeled secondary anti-mouse immunoglobulin antibodies (upper panels) is shown. Staining for α-Gal was a one-step incubation; hence, −lectin on the histogram represents cells only, and +lectin represents cells incubated with IB-4 lectin conjugated to FITC (lower panels). (b) α-Gal expression on ST-IOWA wild-type and ST-IOWA Gal-null (−/−) cells. Expression of the α-Gal antigen was analyzed by FACs, using the IB-4 lectin. A rightward shift of the histogram in the presence of lectin (+lectin) compared to the histogram generated in the absence of lectin (−lectin) indicates expression of the α-Gal antigen. The level of expression was determined using the MFI shift, generated by the Cell Quest FACScan software. The MFI shift was calculated by determining test MFI and subtracting that of negative controls (no primary antibodies for hCD55 and hCD46 expression and no lectin for α-Gal expression), and all the mean shifts are shown in Table ​Table11.

Article Snippet: Samples were then incubated on ice with either primary mouse anti-human CD55 antibody (BRIC110) or mouse anti-human CD46 antibody (J4-48) from Cymbus Bioscience Ltd., diluted to 1:20 in PBS/BA, for 1 h. After three washes in PBS/BA, samples were incubated with fluorescein isothiocyanate (FITC)-conjugated anti-mouse secondary antibody (Jackson ImmunoResearch), diluted to 1:200 in PBS/BA and incubated on ice for 1 h. For α-Gal expression, cells were detached by a cell scraper, washed, resuspended, and stained in a single 1-h incubation with FITC-conjugated Bandeiraea simplicifolia isolectin (IB-4) (Sigma) at 10 μg/ml in PBS/BA on ice, as this lectin is specific for the terminal α-Gal sugar.

Techniques: Expressing, Transgenic Assay, Staining, Negative Control, Labeling, Incubation, Generated, Software

Summary of virus sensitivity and cell surface molecule expression

Journal:

Article Title: Reduced Sensitivity to Human Serum Inactivation of Enveloped Viruses Produced by Pig Cells Transgenic for Human CD55 or Deficient for the Galactosyl-?(1-3) Galactosyl Epitope

doi: 10.1128/JVI.78.11.5812-5819.2004

Figure Lengend Snippet: Summary of virus sensitivity and cell surface molecule expression

Article Snippet: Samples were then incubated on ice with either primary mouse anti-human CD55 antibody (BRIC110) or mouse anti-human CD46 antibody (J4-48) from Cymbus Bioscience Ltd., diluted to 1:20 in PBS/BA, for 1 h. After three washes in PBS/BA, samples were incubated with fluorescein isothiocyanate (FITC)-conjugated anti-mouse secondary antibody (Jackson ImmunoResearch), diluted to 1:200 in PBS/BA and incubated on ice for 1 h. For α-Gal expression, cells were detached by a cell scraper, washed, resuspended, and stained in a single 1-h incubation with FITC-conjugated Bandeiraea simplicifolia isolectin (IB-4) (Sigma) at 10 μg/ml in PBS/BA on ice, as this lectin is specific for the terminal α-Gal sugar.

Techniques: Expressing

Demonstration of hCD55 incorporation on VSV particles by a viral pull-down assay. VSV harvested through HeLa, TgPAE A, and PAE E cells in the presence of antibody was incubated with protein G-expressing bacterial cells (OMNISORB). Antibodies used were three anti-human CD55 antibodies, BRIC 216, BRIC 471, and anti-DAF; anti-human CD46 J4-48 (hCD46); and anti-CD59 BRA-10G (hCD59). Five micrograms of anti-DAF and 1 μg of the other antibodies were used for incubation with virus and protein G cells. Titers of VSV for pulled-down and input particles were determined by TCID50 assay. The ratio of these titers is shown as percent binding.

Journal:

Article Title: Reduced Sensitivity to Human Serum Inactivation of Enveloped Viruses Produced by Pig Cells Transgenic for Human CD55 or Deficient for the Galactosyl-?(1-3) Galactosyl Epitope

doi: 10.1128/JVI.78.11.5812-5819.2004

Figure Lengend Snippet: Demonstration of hCD55 incorporation on VSV particles by a viral pull-down assay. VSV harvested through HeLa, TgPAE A, and PAE E cells in the presence of antibody was incubated with protein G-expressing bacterial cells (OMNISORB). Antibodies used were three anti-human CD55 antibodies, BRIC 216, BRIC 471, and anti-DAF; anti-human CD46 J4-48 (hCD46); and anti-CD59 BRA-10G (hCD59). Five micrograms of anti-DAF and 1 μg of the other antibodies were used for incubation with virus and protein G cells. Titers of VSV for pulled-down and input particles were determined by TCID50 assay. The ratio of these titers is shown as percent binding.

Article Snippet: Samples were then incubated on ice with either primary mouse anti-human CD55 antibody (BRIC110) or mouse anti-human CD46 antibody (J4-48) from Cymbus Bioscience Ltd., diluted to 1:20 in PBS/BA, for 1 h. After three washes in PBS/BA, samples were incubated with fluorescein isothiocyanate (FITC)-conjugated anti-mouse secondary antibody (Jackson ImmunoResearch), diluted to 1:200 in PBS/BA and incubated on ice for 1 h. For α-Gal expression, cells were detached by a cell scraper, washed, resuspended, and stained in a single 1-h incubation with FITC-conjugated Bandeiraea simplicifolia isolectin (IB-4) (Sigma) at 10 μg/ml in PBS/BA on ice, as this lectin is specific for the terminal α-Gal sugar.

Techniques: Pull Down Assay, Incubation, Expressing, TCID50 Assay, Binding Assay