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complement elisa kits microview ba  (Quidel)
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Quidel complement elisa kits microview ba
(A) NHS was incubated with 50 μM hemin, 12.5 μM Hb (1 molecule of Hb contains 4 heme molecules) diluted in TBS. After a 30-minute incubation time at 37°C, the level of released Ba was measured by <t>ELISA.</t> *P < 0.005, Kruskal-Wallis with Dunn’s test for multiple pairwise comparisons. (B) HUVECs were treated with increasing concentrations of hemin or diluted in FCS-free culture medium (M199) for 30 minutes at 37°C. After removing the supernatant, HUVECs were exposed to NHS (33% final concentration diluted in the same medium) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition by flow cytometry. (C–F) HUVECs were treated with increased concentrations of hemin with or without 5 μM of Hx for 30 minutes at 37°C. After removing the supernatant, HUVECs were exposed to NHS (33% final concentration diluted in M199 FCS-free) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition (C and D) or C5b-9 formation (E and F) by flow cytometry. (C and E) Representative flow cytometry histograms. (D and F) Quantitative analyses. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, 2-way ANOVA with Sidak’s test for multiple comparisons. Values are shown as box plots with median and minimum/maximum points.
Complement Elisa Kits Microview Ba, supplied by Quidel, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) NHS was incubated with 50 μM hemin, 12.5 μM Hb (1 molecule of Hb contains 4 heme molecules) diluted in TBS. After a 30-minute incubation time at 37°C, the level of released Ba was measured by ELISA. *P < 0.005, Kruskal-Wallis with Dunn’s test for multiple pairwise comparisons. (B) HUVECs were treated with increasing concentrations of hemin or diluted in FCS-free culture medium (M199) for 30 minutes at 37°C. After removing the supernatant, HUVECs were exposed to NHS (33% final concentration diluted in the same medium) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition by flow cytometry. (C–F) HUVECs were treated with increased concentrations of hemin with or without 5 μM of Hx for 30 minutes at 37°C. After removing the supernatant, HUVECs were exposed to NHS (33% final concentration diluted in M199 FCS-free) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition (C and D) or C5b-9 formation (E and F) by flow cytometry. (C and E) Representative flow cytometry histograms. (D and F) Quantitative analyses. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, 2-way ANOVA with Sidak’s test for multiple comparisons. Values are shown as box plots with median and minimum/maximum points.

Journal: JCI Insight

Article Title: Intravascular hemolysis activates complement via cell-free heme and heme-loaded microvesicles

doi: 10.1172/jci.insight.96910

Figure Lengend Snippet: (A) NHS was incubated with 50 μM hemin, 12.5 μM Hb (1 molecule of Hb contains 4 heme molecules) diluted in TBS. After a 30-minute incubation time at 37°C, the level of released Ba was measured by ELISA. *P < 0.005, Kruskal-Wallis with Dunn’s test for multiple pairwise comparisons. (B) HUVECs were treated with increasing concentrations of hemin or diluted in FCS-free culture medium (M199) for 30 minutes at 37°C. After removing the supernatant, HUVECs were exposed to NHS (33% final concentration diluted in the same medium) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition by flow cytometry. (C–F) HUVECs were treated with increased concentrations of hemin with or without 5 μM of Hx for 30 minutes at 37°C. After removing the supernatant, HUVECs were exposed to NHS (33% final concentration diluted in M199 FCS-free) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition (C and D) or C5b-9 formation (E and F) by flow cytometry. (C and E) Representative flow cytometry histograms. (D and F) Quantitative analyses. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, 2-way ANOVA with Sidak’s test for multiple comparisons. Values are shown as box plots with median and minimum/maximum points.

Article Snippet: Complement ELISA kits MicroView Ba and sC5b-9 were from Quidel.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Staining, Flow Cytometry

(A and B) NHS was incubated with 1,000 MV/μl RBC MVs from HDs (n = 12) or SCD patients (n = 12) diluted in TBS. After 30 minutes at 37°C, the levels of released Ba (A) and sC5b-9 (B) were measured by ELISA. **P < 0.005, Mann-Whitney test. (C) Linear correlation between measured Ba and sC5b-9 levels with RBC MPs from SCD (n = 12) in tested samples. R2 = 0.4811; P = 0.0085. (D and E) NHS was incubated with 1,000 MV/μl MVs from SCD patients (n = 12) diluted in TBS with or without 25 μM Hx. After 30 minutes at 37°C, the levels of released Ba (D) and sC5b-9 (E) were measured by ELISA. *P < 0.05, ***P < 0.001, Wilcoxon test after a Shapiro-Wilk test for normality. (F and G) HUVECs were treated with 1,000 MV/μl RBC MPs from HDs (n = 4) or SCD patient (n = 7) diluted in M199 medium for 30 minutes at 37°C. Without removing the supernatant, HUVECs were exposed to NHS (final concentration at 33% diluted in M199 FCS-free) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition by flow cytometry. (F) Flow cytometry histograms. (G) Quantitative analyses. *P < 0.05, Mann-Whitney test. (H) HUVECs were treated with 1,000 MV/μl RBC MPs from 1 SCD patient with or without 5 μM Hx. As above, NHS was added and cells were stained for C3 deposition by flow cytometry. Values are shown as box plots with median and minimum/maximum points.

Journal: JCI Insight

Article Title: Intravascular hemolysis activates complement via cell-free heme and heme-loaded microvesicles

doi: 10.1172/jci.insight.96910

Figure Lengend Snippet: (A and B) NHS was incubated with 1,000 MV/μl RBC MVs from HDs (n = 12) or SCD patients (n = 12) diluted in TBS. After 30 minutes at 37°C, the levels of released Ba (A) and sC5b-9 (B) were measured by ELISA. **P < 0.005, Mann-Whitney test. (C) Linear correlation between measured Ba and sC5b-9 levels with RBC MPs from SCD (n = 12) in tested samples. R2 = 0.4811; P = 0.0085. (D and E) NHS was incubated with 1,000 MV/μl MVs from SCD patients (n = 12) diluted in TBS with or without 25 μM Hx. After 30 minutes at 37°C, the levels of released Ba (D) and sC5b-9 (E) were measured by ELISA. *P < 0.05, ***P < 0.001, Wilcoxon test after a Shapiro-Wilk test for normality. (F and G) HUVECs were treated with 1,000 MV/μl RBC MPs from HDs (n = 4) or SCD patient (n = 7) diluted in M199 medium for 30 minutes at 37°C. Without removing the supernatant, HUVECs were exposed to NHS (final concentration at 33% diluted in M199 FCS-free) for 30 minutes at 37°C. Cells were detached and stained for C3 deposition by flow cytometry. (F) Flow cytometry histograms. (G) Quantitative analyses. *P < 0.05, Mann-Whitney test. (H) HUVECs were treated with 1,000 MV/μl RBC MPs from 1 SCD patient with or without 5 μM Hx. As above, NHS was added and cells were stained for C3 deposition by flow cytometry. Values are shown as box plots with median and minimum/maximum points.

Article Snippet: Complement ELISA kits MicroView Ba and sC5b-9 were from Quidel.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Concentration Assay, Staining, Flow Cytometry