Journal: The FASEB Journal

Article Title: C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease

doi: 10.1096/fj.202200198r

Figure Lengend Snippet: FIGURE 1 The expression of complement effectors C3a, C5a and C5b-9 in peripheral blood and renal biopsies. (A–C) Serum levels of complement C3a, C5a and C5b-9 were examined in healthy controls (n = 20), DM patients (n = 20) and DN patients (n = 20) by ELISA. (D) Representative IHC staining images of complement C3a, C5a and C5b-9 from healthy controls and DN patients. (E) Quantitative analysis of C3a, C5a and C5b-9 staining in renal tissues by Image-Pro Plus 6.0 (n = 10–15). (F) Serum levels of CRP in healthy control (n = 20), DM patients (n = 20) and DN patients (n = 20) detected by ELISA. 400× original magnification; scale bar, 100 µm. Data are presented as the means ± SEM, and p < .05 was considered to be statistically significant

Article Snippet: Usually, the cells were seeded at 1 × 10 5/well in a 12- well plate, and then treated with low glucose (5 mM), high glucose (33 mM), recombinant mouse C3a (100 nM, HY- P7863, MCE, NJ, USA) and recombinant mouse C5a (100 nM, HY- P7695, MCE) for 24 h for qPCR and 72 h for Western blotting.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Control

Journal: The FASEB Journal

Article Title: C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease

doi: 10.1096/fj.202200198r

Figure Lengend Snippet: FIGURE 2 Differences between human CRP and rat CRP in terms of structure and complement activation in vitro. (A) Human CRP and rat CRP migrated in 1/20 SDS-PAGE as pentamers and in SDS-PAGE as monomers. (B) The pentameric structure of human and rat CRP was shown by transmission electron microscopy (TEM). (C) Autologous complement activation of human CRP in human serum and rat CRP in rat serum (n = 6). (D) Interspecific complement activation of human CRP in rat serum and rat CRP in human serum (n = 6). Data are presented as the means ± SEM, and p < .05 was considered to be statistically significant

Article Snippet: Usually, the cells were seeded at 1 × 10 5/well in a 12- well plate, and then treated with low glucose (5 mM), high glucose (33 mM), recombinant mouse C3a (100 nM, HY- P7863, MCE, NJ, USA) and recombinant mouse C5a (100 nM, HY- P7695, MCE) for 24 h for qPCR and 72 h for Western blotting.

Techniques: Activation Assay, In Vitro, SDS Page, Transmission Assay, Electron Microscopy

Journal: The FASEB Journal

Article Title: C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease

doi: 10.1096/fj.202200198r

Figure Lengend Snippet: FIGURE 3 The expression of complement C3a, C5a and C5b-9 in STZ-diabetic DKD rats. (A) Twenty-four-hour microalbuminuria was detected at 14-week with a commercial ELISA kit (n ≥ 8). (B) Blood urea nitrogen, serum CRE and urine CRE were monitored with an automated biochemistry analyzer (n ≥ 8). (C and D) Representative images and quantitative analysis of complement C3a, C5a and C5b-9 in rat kidney tissues by IHC staining (n = 4, more than 20 glomeruli). (E) Serum levels of complement C3a, C5a and C5b-9 were examined in rats with SZT-induced DKD by ELISA (n = 8). 400× original magnification; scale bar, 50 µm. Data are presented as the means ± SEM, and p < .05 was considered to be statistically significant

Article Snippet: Usually, the cells were seeded at 1 × 10 5/well in a 12- well plate, and then treated with low glucose (5 mM), high glucose (33 mM), recombinant mouse C3a (100 nM, HY- P7863, MCE, NJ, USA) and recombinant mouse C5a (100 nM, HY- P7695, MCE) for 24 h for qPCR and 72 h for Western blotting.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

Journal: The FASEB Journal

Article Title: C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease

doi: 10.1096/fj.202200198r

Figure Lengend Snippet: FIGURE 6 Anaphylatoxin C3a induced podocyte autophagy under high-glucose conditions. (A) Quantitative RT–PCR analysis of LC3B and p62 expression under low-glucose and high-glucose conditions with C3a and C5a treatment (n = 3). (B and C) Western blot and quantitative analysis of LC3B and p62 expression under high-glucose conditions with C3a treatment (n = 4). (D) Representative images of autophagy flux changes with C3a and C5a treatment after Ad-mCherry-GFP-LC3B infection (n = 3). 600× original magnification; scale bar, 10 µm. Data are presented as the means ± SEM, p < .05 was considered to be statistically significant

Article Snippet: Usually, the cells were seeded at 1 × 10 5/well in a 12- well plate, and then treated with low glucose (5 mM), high glucose (33 mM), recombinant mouse C3a (100 nM, HY- P7863, MCE, NJ, USA) and recombinant mouse C5a (100 nM, HY- P7695, MCE) for 24 h for qPCR and 72 h for Western blotting.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Infection

Journal: The FASEB Journal

Article Title: C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease

doi: 10.1096/fj.202200198r

Figure Lengend Snippet: FIGURE 7 C3a-induced podocyte autophagy was regulated in stable CRP-overexpression and CRP-knockout cell lines. (A and B) Quantitative RT-PCR analysis of LC3B and p62 expression in the CRP-overexpression experiment and CRP-knockout experiment (n = 3). (C and D) Western blot and quantitative analysis of LC3B and p62 in the CRP-overexpression experiment and CRP-knockout experiment (n = 4). (E and F) Autophagic flux was detected in a stable CRP-overexpression cell line and a stable CRP-knockout cell line under high-glucose conditions with C3a treatment (n = 3). 600× original magnification; scale bar, 10 µm. Data are presented as the means ± SEM, and p < .05 was considered to be statistically significant

Article Snippet: Usually, the cells were seeded at 1 × 10 5/well in a 12- well plate, and then treated with low glucose (5 mM), high glucose (33 mM), recombinant mouse C3a (100 nM, HY- P7863, MCE, NJ, USA) and recombinant mouse C5a (100 nM, HY- P7695, MCE) for 24 h for qPCR and 72 h for Western blotting.

Techniques: Over Expression, Knock-Out, Quantitative RT-PCR, Expressing, Western Blot

Journal: The FASEB Journal

Article Title: C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease

doi: 10.1096/fj.202200198r

Figure Lengend Snippet: FIGURE 8 CRP suppressed C3aR expression under high-glucose conditions. (A) western blot and quantitative analysis of C3aR expression in CRP-overexpression and CRP-knockout experiments (n = 4). (B) IF staining of LC3B in cultured CRP-overexpression and CRP-knockout cell lines with C3a treatment (n = 3). (C) A C3aR agonist active peptide was used to reverse changes in LC3B and p62 expression in CRP-FL cells (n = 4). (D) A C3aR antagonist SB290157 was used to reverse changes in LC3B and p62 expression in CRP- KO2 cells (n = 4). 200× original magnification; scale bar, 10 µm. Data are presented as the means ± SEM, and p < .05 was considered to be statistically significant

Article Snippet: Usually, the cells were seeded at 1 × 10 5/well in a 12- well plate, and then treated with low glucose (5 mM), high glucose (33 mM), recombinant mouse C3a (100 nM, HY- P7863, MCE, NJ, USA) and recombinant mouse C5a (100 nM, HY- P7695, MCE) for 24 h for qPCR and 72 h for Western blotting.

Techniques: Expressing, Western Blot, Over Expression, Knock-Out, Staining, Cell Culture

Journal: Frontiers in Immunology

Article Title: Human Neutrophils Respond to Complement Activation and Inhibition in Microfluidic Devices

doi: 10.3389/fimmu.2021.777932

Figure Lengend Snippet: Microfluidic devices for measuring neutrophil chemotaxis, phagocytosis, and swarming behaviors. (A) Microscopic image of a segment of the chemotaxis microfluidic device (x10 Brightfield, Nikon TiE) showing neutrophil chemotaxis towards C5a and C3a during increasing mechanical restriction. Each chemoattractant chamber is connected to the cell loading channel through 3 tapered channels. (B) Microscopic image of a segment of the phagocytosis microfluidic device (x10 Brightfield/Fluorescent, Nikon TiE) showing neutrophils (blue nuclei) from the outer chamber migrating towards the central reservoir through a migration channel to phagocytose S. aureus particles (green) in plasma at T 20 minutes. (C) Microscopic images obtained at three different time points (T 0 , T 10 , and T 30 minutes) showing the formation of a neutrophil swarm (blue nuclei) over a cluster of Texas red-labeled zymosan A S. cerevisiae bioparticles (Red).

Article Snippet: The chemoattractant chambers were filled either with fMLP (N-formyl-methionyl-leucyl-phenylalanine, 100 nM, Millipore Sigma), recombinant human C5a protein (ab61918, 0.1 - 10 μM, Abcam), or C3a (3677-C3-025, 0.1 - 10 μM, R&D Systems) diluted in HBSS with 0.5% BSA during the initial priming step.

Techniques: Chemotaxis Assay, Migration, Clinical Proteomics, Labeling

Journal: Frontiers in Immunology

Article Title: Human Neutrophils Respond to Complement Activation and Inhibition in Microfluidic Devices

doi: 10.3389/fimmu.2021.777932

Figure Lengend Snippet: Chemotaxis of isolated neutrophils towards C5a and C3a and the effect of AVA on chemotaxis. (A) The percentage of neutrophils migrating directionally towards increasing concentrations of C5a and C3a compared to media alone (HBSS with 0.5%BSA, N=3, ****p < 0.0001; One-way ANOVA with Dunnett’s test) (B) C3a inhibits neutrophil chemotaxis towards C5a, C5a effect is dependent on concentration, and there is no interaction effect between C3 and C5a. (N=3 donors, p < 0.005, Repeated measures, two way ANOVA, with Sidak’s correction for multiple comparisons). (C) Percentage of neutrophils migrating directionally towards C5a (100 nM) after treatment of neutrophils with C5aR1 antagonist AVA (N=3, 4 or 5, each dot on the plot represents one experiment, ****p < 0.01 for comparisons to C5a control; ns represents not statistical significant. One way ANOVA with Dunnett’s multiple comparison test with single pooled variance).

Article Snippet: The chemoattractant chambers were filled either with fMLP (N-formyl-methionyl-leucyl-phenylalanine, 100 nM, Millipore Sigma), recombinant human C5a protein (ab61918, 0.1 - 10 μM, Abcam), or C3a (3677-C3-025, 0.1 - 10 μM, R&D Systems) diluted in HBSS with 0.5% BSA during the initial priming step.

Techniques: Chemotaxis Assay, Isolation, Concentration Assay, Control, Comparison

Journal: Frontiers in Immunology

Article Title: Human Neutrophils Respond to Complement Activation and Inhibition in Microfluidic Devices

doi: 10.3389/fimmu.2021.777932

Figure Lengend Snippet: Atypical membrane elasticity of neutrophils and migration patterns when exposed to high C5a dose. (A) Percentage of neutrophil migration patterns during chemotaxis towards C5a, C3a, and fMLP. (N=3). (B) Microscopic images of elongated neutrophils when exposed to C5a 1µM inside the end chambers. Scale = 100 µm. (C) Percentages of unique reversed migration phenotype of neutrophils observed when exposed to high C5a concentration. (N=3, *p < 0.05; ns represents not statistical significant. One-way ANOVA with Dunnett’s test).

Article Snippet: The chemoattractant chambers were filled either with fMLP (N-formyl-methionyl-leucyl-phenylalanine, 100 nM, Millipore Sigma), recombinant human C5a protein (ab61918, 0.1 - 10 μM, Abcam), or C3a (3677-C3-025, 0.1 - 10 μM, R&D Systems) diluted in HBSS with 0.5% BSA during the initial priming step.

Techniques: Membrane, Migration, Chemotaxis Assay, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Human Neutrophils Respond to Complement Activation and Inhibition in Microfluidic Devices

doi: 10.3389/fimmu.2021.777932

Figure Lengend Snippet: Effect of C5 complement inhibitors on neutrophil chemotaxis towards endogenous complement activation with CVF. (A) Effect of AVA (250 nM) on endogenous complement activation by increasing CVF doses (N=3 donors, *p < 0.05, **p < 0.01; paired two-tailed t -test). (B) Effect of C5 inhibition with ECU (3 µM) and RA101295 (3 µM) on endogenous complement activation by CVF (N=3, **p < 0.01; ns represents not statistical significant. One-way ANOVA with Dunnett’s test).

Article Snippet: The chemoattractant chambers were filled either with fMLP (N-formyl-methionyl-leucyl-phenylalanine, 100 nM, Millipore Sigma), recombinant human C5a protein (ab61918, 0.1 - 10 μM, Abcam), or C3a (3677-C3-025, 0.1 - 10 μM, R&D Systems) diluted in HBSS with 0.5% BSA during the initial priming step.

Techniques: Chemotaxis Assay, Activation Assay, Two Tailed Test, Inhibition

Journal: Frontiers in Immunology

Article Title: Human Neutrophils Respond to Complement Activation and Inhibition in Microfluidic Devices

doi: 10.3389/fimmu.2021.777932

Figure Lengend Snippet: Schematic representation of complement activation pathways, terminal inhibition strategies, and their effects on neutrophil functional behaviors. The classical, lectin, and alternative pathways converge on the cleavage of the central components C3 into C3a and C3b peptides. C3a acts as an anti-inflammatory mediator towards neutrophils, while C3b enhances phagocytosis through opsonization and forms C5 convertases, which cleaves C5 into C5a and C5b. C5a is a potent pro-inflammatory mediator and chemoattractant, while C5b forms the membrane attack complex (MAC), leading to cell and bacteria lysis. ECU and RA101295 are C5-inhibitors, which block the production of C5a and C5b peptides, while AVA blocks C5a mediated chemotaxis and pro-inflammatory effects.

Article Snippet: The chemoattractant chambers were filled either with fMLP (N-formyl-methionyl-leucyl-phenylalanine, 100 nM, Millipore Sigma), recombinant human C5a protein (ab61918, 0.1 - 10 μM, Abcam), or C3a (3677-C3-025, 0.1 - 10 μM, R&D Systems) diluted in HBSS with 0.5% BSA during the initial priming step.

Techniques: Activation Assay, Inhibition, Functional Assay, Membrane, Bacteria, Lysis, Blocking Assay, Chemotaxis Assay

Journal: Mucosal immunology

Article Title: Activation of C3a receptor is required in cigarette smoke-mediated emphysema

doi: 10.1038/mi.2014.118

Figure Lengend Snippet: Purified human C3 (100μg/ml) was cleaved with different concentrations of human ( a ) NE (10μg/ml, 1μg/ml, 0.1μg/ml) and ( b ) MMP12 (10μg/ml, 1μg/ml, 0.1μg/ml) for 4 hours at 37°C. Cleavage products were separated using 10% non-reducing Tricine gels, and detected by Western blot using anti-C3a antibody; purified C3a, NE and MMP12 were loaded as controls. ( c ) Bone marrow-derived dendritic cells (BMDCs; 5×10 4 ) and ( d ) myeloid-derived dendritic cells (MDDCs; 5×10 4 ) were suspended in media (RPMI-1640) and were placed on 48-well chemotaxis chambers for 1hr in the presence of intact or MMP12, NE cleaved C3 protein; control conditions included NE and MMP12. Transmigrating cells were detected in stained membranes visualized under microscope (20x) and reported as the average number of cells/field (n=4-6). **P<0.01, as determined by one-way ANOVA with Bonferroni's multiple comparison. Results are represented as mean±s.e.m, from 3 independent experiments.

Article Snippet: Antibodies used for western blotting and immunohistochemistry of C3 and monoclonal anti-human complement component C3a antibody were purchased from R&D Systems (Minneapolis, MN); mouse anti-human complement C3b-alpha monoclonal antibody for detection of C3b and C3c was purchased from Chemicon, Millipore (Billerica, MA); mouse anti-human C3d were purchased from AbD Serotec (Raleigh, NC).

Techniques: Purification, Western Blot, Derivative Assay, Chemotaxis Assay, Control, Staining, Microscopy, Comparison

Journal: Mucosal immunology

Article Title: Activation of C3a receptor is required in cigarette smoke-mediated emphysema

doi: 10.1038/mi.2014.118

Figure Lengend Snippet: WT and C3 deficient mice were exposed to cigarette smoke or air for 6 months. ( a ) Expression of C3ar1 mRNA in BAL cells isolated from WT or C3 −/− mice exposed to air or cigarette smoke was measured by quantitative reverse transcription PCR (qPCR). ***P<0.001, as determined by the one-way ANOVA with Bonferroni's multiple comparison. Representative ( b ) and cumulative data ( c ) measuring C3aR mean fluorescent intensity (MFI) in single lung cells gated on B220 − CD11c + population using flow cytometry. *P<0.05, as determined by the one-way ANOVA with Bonferroni's multiple comparison. ( d ) Representative photomicrograph of WT and C3 −/− mouse lung tissue exposed to six months of smoke or air immunostained for expression of C3aR (green) or nuclei (blue; DAPI). Scale bar: 20μm. Green arrows indicate C3aR + cells. ( e ) to ( h ) Mouse bone marrow-derived dendritic cells (BMDCs; 2×10 5 ) were treated with C3aR Agonist (CAS 944997-60-8; 20ng/ml) or vehicle (2% DMSO) for 48 hours. Expression level of C3aR1, Il6, Mmp9 and Mmp12 mRNA were measured using qPCR (n=4 in each group; **P<0.01, ***P<0.001, as determined by student t test. ( i ) Human CD1a + lung mDCs (2×10 5 ) were treated with purified human C3a (40ng/ml) for 24 hours or vehicle (media). Expression level of C3AR1 mRNA was measured by quantitative reverse transcription PCR (qPCR). n=3; **P<0.01, as determined by student t test. All gene expressions were normalized to 18S ribosomal RNA expression and analyzed by ΔΔCt . Results are represented as mean±s.e.m, from 3 independent experiments with 4-5 mice in each group ( a-d ).

Article Snippet: Antibodies used for western blotting and immunohistochemistry of C3 and monoclonal anti-human complement component C3a antibody were purchased from R&D Systems (Minneapolis, MN); mouse anti-human complement C3b-alpha monoclonal antibody for detection of C3b and C3c was purchased from Chemicon, Millipore (Billerica, MA); mouse anti-human C3d were purchased from AbD Serotec (Raleigh, NC).

Techniques: Expressing, Isolation, Reverse Transcription, Comparison, Flow Cytometry, Derivative Assay, Purification, RNA Expression

Journal: Journal of Alzheimer's Disease

Article Title: Impact of Preanalytical Procedures on Complement Biomarkers in Cerebrospinal Fluid and Plasma from Controls and Alzheimer’s Disease Patients

doi: 10.3233/JAD-240287

Figure Lengend Snippet: Sensitivity, precision and specificity results for the complement assays

Article Snippet: The C3 assay measures full-length C3 using anti-human C3a polyclonal antibody (R& D Systems, catalog AF3677) as a capture antibody and polyclonal anti-human C3 antibody for detection (MP Bio, catalog 0855033).

Techniques:

Journal: Journal of Alzheimer's Disease

Article Title: Impact of Preanalytical Procedures on Complement Biomarkers in Cerebrospinal Fluid and Plasma from Controls and Alzheimer’s Disease Patients

doi: 10.3233/JAD-240287

Figure Lengend Snippet: (a) Schematic of the study design to assess complement stability in plasma or CSF from HC and AD subjects. CSF and blood were collected from 9 donors, the biofluids were centrifuged and aliquoted at the clinical site without (No Addition) or with supplementation of the complement-stabilizing agents EDTA, EDTA + FUT and PIC. The CSF and plasma aliquots underwent 1-4 FT cycles. C4a, C3a, Bb, C4, C3, and FB concentrations were assessed by immunoassay. Created with BioRender.com. (b) AD diagnostic biomarkers Aβ 42 and Aβ 40 were measured by immunoassay in non-supplemented CSF samples from each donor. Aβ 42 :Aβ 40 ratio (AU) in HC (amyloid negative, N = 4) and AD (amyloid positive, N = 5) subjects; horizontal and vertical lines represent the mean and SD respectively. The dashed line represents the diagnostic cutoff value of 0.055.

Article Snippet: The C3 assay measures full-length C3 using anti-human C3a polyclonal antibody (R& D Systems, catalog AF3677) as a capture antibody and polyclonal anti-human C3 antibody for detection (MP Bio, catalog 0855033).

Techniques: Diagnostic Assay

Journal: Journal of Alzheimer's Disease

Article Title: Impact of Preanalytical Procedures on Complement Biomarkers in Cerebrospinal Fluid and Plasma from Controls and Alzheimer’s Disease Patients

doi: 10.3233/JAD-240287

Figure Lengend Snippet: (a–b) Impact of the different additives on complement levels in plasma (a) and CSF (b). Shown are the % recovery from the unsupplemented condition (No Addition) in the supplemented conditions (EDTA, EDTA + FUT, and PIC) for C4a, C3a, Bb, C4, C3, and FB at FT1. The dashed line represents the ‘No Addition’ reference (100%). Box plots shown comprise data from N = 9 individual donors. Boxes represent the median and interquartile range (IQR); The lower and upper hinges correspond to the first and third quartiles (the 25th and 75th percentiles). The upper whisker extends from the hinge to the largest value no further than 1.5 * IQR from the hinge. The lower whisker extends from the hinge to the smallest value at most 1.5 * IQR of the hinge. The diamond shape represents the mean. Individual data points are plotted over the box plots.

Article Snippet: The C3 assay measures full-length C3 using anti-human C3a polyclonal antibody (R& D Systems, catalog AF3677) as a capture antibody and polyclonal anti-human C3 antibody for detection (MP Bio, catalog 0855033).

Techniques: Whisker Assay

Journal: Journal of Alzheimer's Disease

Article Title: Impact of Preanalytical Procedures on Complement Biomarkers in Cerebrospinal Fluid and Plasma from Controls and Alzheimer’s Disease Patients

doi: 10.3233/JAD-240287

Figure Lengend Snippet: Impact of FT cycles on complement levels in plasma. Shown are the % recovery from FT1 in the FT2, FT3, and FT4 unsupplemented (No Addition) or supplemented aliquots (EDTA, EDTA + FUT, PIC) for C4a, C3a, Bb, C4, C3, and FB. The dashed line represents the ‘FT1’ reference (100%). Box plots shown comprise data from N = 9 individual donors. Boxes represent the median and interquartile range (IQR); The lower and upper hinges correspond to the first and third quartiles (the 25th and 75th percentiles). The upper whisker extends from the hinge to the largest value no further than 1.5 * IQR from the hinge. The lower whisker extends from the hinge to the smallest value at most 1.5 * IQR of the hinge. The diamond shape represents the mean. Individual data points are plotted over the box plots.

Article Snippet: The C3 assay measures full-length C3 using anti-human C3a polyclonal antibody (R& D Systems, catalog AF3677) as a capture antibody and polyclonal anti-human C3 antibody for detection (MP Bio, catalog 0855033).

Techniques: Whisker Assay

Journal: Journal of Alzheimer's Disease

Article Title: Impact of Preanalytical Procedures on Complement Biomarkers in Cerebrospinal Fluid and Plasma from Controls and Alzheimer’s Disease Patients

doi: 10.3233/JAD-240287

Figure Lengend Snippet: Impact of FT cycles on complement levels in CSF. Shown are the % recovery from FT1 in the FT2, FT3 and FT4 unsupplemented (No Addition) or supplemented aliquots (EDTA, EDTA + FUT, PIC) for C4a, C3a, Bb, C4, C3, and FB. The dashed line represents the ‘FT1’ reference (100%). Box plots shown comprise data from N = 9 individual donors. Boxes represent the median and interquartile range (IQR); The lower and upper hinges correspond to the first and third quartiles (the 25th and 75th percentiles). The upper whisker extends from the hinge to the largest value no further than 1.5 * IQR from the hinge. The lower whisker extends from the hinge to the smallest value at most 1.5 * IQR of the hinge. The diamond shape represents the mean. Individual data points are plotted over the box plots.

Article Snippet: The C3 assay measures full-length C3 using anti-human C3a polyclonal antibody (R& D Systems, catalog AF3677) as a capture antibody and polyclonal anti-human C3 antibody for detection (MP Bio, catalog 0855033).

Techniques: Whisker Assay

Journal: Journal of Alzheimer's Disease

Article Title: Impact of Preanalytical Procedures on Complement Biomarkers in Cerebrospinal Fluid and Plasma from Controls and Alzheimer’s Disease Patients

doi: 10.3233/JAD-240287

Figure Lengend Snippet: (a–b) Rank order preservation of complement levels between the first (FT1) and second (FT2) freeze-thaw cycle in plasma (a) and CSF (b). Lollipop plot showing the Spearman Correlation Coefficient between the concentrations at FT1 and the concentrations at FT2 in unsupplemented (No Addition) or supplemented aliquots (EDTA, EDTA + FUT, PIC) for C4a, C3a, Bb, C4, C3, and FB.

Article Snippet: The C3 assay measures full-length C3 using anti-human C3a polyclonal antibody (R& D Systems, catalog AF3677) as a capture antibody and polyclonal anti-human C3 antibody for detection (MP Bio, catalog 0855033).

Techniques: Preserving