goat anti-factor b polyclonal antibody  (Complement Technology Inc)

Journal: bioRxiv

Article Title: Mucosally sourced complement factor B modulates the host response to colitis

doi: 10.64898/2025.12.31.697141

Figure Lengend Snippet: ( A ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway in intestinal tissue biopsies from uninflamed and non-inflamed regions of patients with ulcerative colitis (UC, Single Cell Portal: SCP259). ( B ) Plots depict CFB expression across denoted colonic cell types in the same dataset from intestinal tissue of patients with UC and healthy controls. * P <0.05, *** P <0.001, ns, not significant, Wilcoxon rank test. ( C ) Immunofluorescence staining for Factor B (red), E-cadherin (green) and DAPI (blue) in colonic tissue specimens from patients with IBD versus controls (non-IBD). (Scale bar = 90 μ m). ( D ) Quantification of the relative intensity of Factor B expression across tissue type. ( E ) Representative spatial transcriptomic slides (Visium) showing CFB expression in intestinal tissues obtained from patients with UC compared to healthy controls (HC, GSE189184). Graph depicts quantification of CFB expression from 3 HC and 7 UC specimens. ( F ) Representative spatial transcriptomic slides showing DUOX2-CFB co-localization spots in HC (left) and UC tissue (right) from the same dataset. Bar plot represents average co-expression across tissue samples. Error bars represent standard error of the mean (SEM). * P < 0.05, unpaired t-test.

Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Mucosally sourced complement factor B modulates the host response to colitis

doi: 10.64898/2025.12.31.697141

Figure Lengend Snippet: ( A ) Immunoblot denoting Factor B (FB) in plasma of global Cfb knockouts ( Cfb −/− ), mice deficient in liver-derived FB ( Cfb f/f AlbCre +/− ) and their littermates ( Cfb f/f AlbCre −/− ). MM: molecular marker. The same volume of plasma was loaded per well at 1:20 dilution. ( B ) Cfb expression on qRT-PCR in the colons from indicated mice, values normalized to Hprt1 . ( C ) Quantification of the relative intensity of Factor B expression in colon tissue from indicated mice. ( D ) Representative immunofluorescence images of colonic tissue from Cfb −/− , Cfb f/f AlbCre +/− and Cfb f/ f AlbCre −/− mice (n=4/group), stained with Factor B (red), E-cadherin (green) and DAPI (blue). Scale bars = 70 μ m. ( E ) Graph depicts serum CD14 concentration. Serum from mice challenged with lipopolysaccharide (LPS) were used as a positive control for barrier leak. Each dot represents an individual mouse. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant, unpaired t test. ( F ) Dot plot illustrating the expression of complement genes representing the classical (CP), lectin (LP) and alternative (AP) pathway across cell types indicated along the Y axis, based on single-cell RNA-seq data from the large intestines of non-injured adult C57BL/6 mice ( Cfb f/f ), housed in specific pathogen-free conditions. Cells pooled from n=2 mice.

Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

Techniques: Western Blot, Clinical Proteomics, Derivative Assay, Marker, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Concentration Assay, Positive Control, RNA Sequencing

Journal: bioRxiv

Article Title: Mucosally sourced complement factor B modulates the host response to colitis

doi: 10.64898/2025.12.31.697141

Figure Lengend Snippet: ( A-B ) Human epithelial Caco-2 cells were incubated in serum-free media alone (non-treated, NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). ( A ) Supernatant was collected after 24 hours, concentrated 50-fold and analyzed by immunoblotting for Factor B. ( B ) Human primary fibroblasts were incubated in serum-free media alone (NT), media with rIL-1β (50ng/ml) or TNF-α (50ng/ml). Immunoblot depicts CFB in supernatant at the 24 h time point. Graph depicts Factor B protein quantification relative to the loaded control. The same volume of supernatant was loaded in each well. Each dot represents an individual replicate. Bar graphs show the mean ± SEM. All immunoblots were repeated at least twice. Statistics performed using an ordinary one-way ANOVA test adjusted for multiple comparisons.

Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

Techniques: Incubation, Western Blot, Control

Journal: bioRxiv

Article Title: Mucosally sourced complement factor B modulates the host response to colitis

doi: 10.64898/2025.12.31.697141

Figure Lengend Snippet: ( A ) SPF-housed global knockout ( Cfb −/− ), liver-deficient ( Cfb f/f AlbCre +/− ) and littermate control ( Cfb f/f AlbCre −/− ) mice were administered 2.5% DSS in drinking water for 7 days, followed by conventional water for 3 days. Mice were euthanized on Day 11. Body weights were monitored daily during treatment. ( B ) Colon lengths evaluated at end of study. ( C ) Colonic tissue was homogenized for measuring TNF-α and MPO levels, normalized to weight (per g) of tissue. ( D ) Fecal lipocalin-2 (LCN-2) levels compared at the end of the study. ( E ) Complement activity in the colonic tissue measured using C3bBbP (alternative pathway convertase). ( F ) Differentially regulated pathways upregulated in Cfb −/− versus Cfb f/f AlbCre +/− based on bulk RNA-seq of colonic tissue at end of treatment. Pathway analysis conducted using Enrichr (MSigDB). ( G ) Adult C57BL/6 mice housed in SPF conditions were given 2.5% DSS in drinking water for a week. Starting day 5, mice were treated with a daily oral gavage of a Factor B inhibitor, iptacopan (20 mg/kg), or vehicle control. Body weights were monitored daily. ( H ) Colon lengths compared at end of the experiment. ( I ) IL-1β in colonic tissue lysate and ( J ) LCN-2 in fecal samples. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

Techniques: Knock-Out, Control, Activity Assay, RNA Sequencing, Comparison

Journal: bioRxiv

Article Title: Mucosally sourced complement factor B modulates the host response to colitis

doi: 10.64898/2025.12.31.697141

Figure Lengend Snippet: ( A-C ) Data from scRNA-seq of colonic cells from C57BL/6 mice subjected to acute DSS (acute colitis, AC) was re-analyzed for express of complement genes and compared to mice not treated with DSS (non-DSS, healthy controls (HC), GSE264408). ( A ) UMAP with designated cell clusters showing Cfb expression. ( B ) Bar plots depicting changes in the cellular composition of the colonic tissue post-DSS treatment. ( C ) Dot plots showing average gene expression levels of key AP components in the distinct cell clusters of HC (healthy control) mice and those with AC (acute colitis). ( D ) Cfb f/f LysMCre +/− mice and controls were treated with 7 days of 2.5% DSS in drinking water, followed by 3 days of regular water. Body weights were monitored daily. Colon length was determined at the end of the experiment. Colonic tissue was homogenized for comparing MPO and IL-1β levels, normalized to tissue weight. ( E ) Representative RNAScope images depict staining of colonic tissue from Cfb f/f VilCre +/− mice and controls. Scale bar =30μm. ( F ) Cfb f/f VilCre and controls were treated similarly to ( D ). Body weights and the disease activity index (DAI) were measured daily. Mice were sacrificed on day 11. Colons were harvested to compare the colon length, MPO and IL-1β levels, normalized to weight of tissue. ( G ) Cfb f/f Col1a2CreER T2+/− mice and their controls were treated with tamoxifen every other day for 10 days. Colons were harvested and tissue sections were stained for Factor B (red) and vimentin (green). Scale bar =90 μ m. ( H ) Cfb f/f Col1a2CreER T2+/− and littermates were subjected to tamoxifen challenge as detailed in ( G ). 3 days after the last dose of tamoxifen, mice were subjected to DSS challenge as detailed in ( D ). Body weights and DAI scores were monitored daily. Colon tissue was collected at day 11 to measure colon length and cytokines in tissue lysate. Data represent 2 independent experiments. Bar graphs show the mean ± SEM, * P < 0.05; ** P < 0.01, ns, non-significant. Unpaired t test when analyzing a two-group categorical comparison, and 2-way ANOVA adjusted for multiple comparisons when analyzing longitudinal data.

Article Snippet: The sections were then blocked with 3% Bovine Albumen (Thermo Fisher, #J10857.22) diluted in phosphate buffered saline (PBS) at room temperature for 1 h. For primary incubation the sections were stained with a 1:50 dilution of polyclonal rabbit anti-goat Factor B antibody (Quidel Ortho, # A311) and a 1:100 dilution of mouse anti-E-Cadherin (BD Sciences, Cat. 610181) overnight at 4°C.

Techniques: Expressing, Gene Expression, Control, RNAscope, Staining, Activity Assay, Comparison

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Proteins’ Western Blotting Information.

Article Snippet: 6 , CFB , 85 kDa , Goat Anti-factor B Polyclonal antibody # AF2739 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Simvastatin attenuates radiation-induced salivary gland dysfunction in mice

doi: 10.2147/dddt.s105809

Figure Lengend Snippet: Figure 5 Protein content of TGF-β1 (25 Kd) in SMGs of mice at 8 hours, 24 hours, and 4 weeks after IR was assessed by Western blot analysis. Notes: (A) Representative Western blots from three to five experiments with similar results. (B) Changes in TGF-β1 quantified by scanning densitometry analysis using Image Lab software. The data (relative density normalized to β-actin) are expressed as mean ± SD; *P,0.05, **P,0.01. Abbreviations: TGF-β1, transforming growth factor β1; SMG, submandibular gland; IR, irradiation; SD, standard deviation; h, hours; w, weeks.

Article Snippet: The membranes were blocked in a blocking buffer (5% nonfat milk, 1% Tween 20, in 20 mM Tris-buffered saline, pH 7.6) for 1 hour at room temperature, followed by incubation with the transforming growth factor (TGF)-β1 rabbit polyclonal immunoglobulin G (IgG) (Santa Cruz Biotechnology Inc., Dallas, TX, USA) in a blocking buffer overnight at 4°C.

Techniques: Western Blot, Software, Irradiation, Standard Deviation

Journal: PloS one

Article Title: Effects of angiopoietin-1 on hemorrhagic transformation and cerebral edema after tissue plasminogen activator treatment for ischemic stroke in rats.

doi: 10.1371/journal.pone.0098639

Figure Lengend Snippet: Figure 1. Endogenous Ang1 localization in non-ischemic rat brains visualized with confocal laser microscopy. (A) From left to right: markers of cells that make up the blood brain barrier (a, e, i, m; green), Ang1 (b, f, j, n; red), 49,6-diamidino-2-phenylindole (DAPI) stain (c, g, k, o; blue), and a merged image (d, h, l, p). RECA1 is an endothelial cell marker protein; PDGFRb is a pericyte marker; GFAP is an astrocyte marker; and MAP2 is a neuronal cell marker. RECA1, rat endothelial cell antigen; PDGFRb, platelet-derived growth factor receptor b; GFAP, glial fibrillary acidic protein; MAP2, microtubule-associated protein 2; Ang1, angiopoietin-1. The scale bars are 10 mm. (B) Three-dimensional (3D) images of endogenous Ang1 in non- ischemic brains visualized with a confocal laser microscope. a: endothelial cell marker RECA1 (green) and endogenous Ang1 (red); b: neuronal cell marker MAP2 (green) and endogenous Ang1 (red). RECA1, rat endothelial cell antigen; Ang1, angiopoietin-1; MAP2, microtubule-associated protein 2. An arrow indicates Ang1 expressed in a pericyte. Arrowheads indicate Ang1 expressed in neuronal cells. doi:10.1371/journal.pone.0098639.g001

Article Snippet: The primary antibodies that were used were a rabbit polyclonal anti-Ang1 antibody (ab93599: Abcam plc, 1:100), mouse monoclonal anti-RECA1 antibody (MCA-970R: AbD Serotec, 1:250), mouse monoclonal anti-glial fibrillary acidic protein (GFAP) antibody (3670: Cell Signaling Technology, Inc., 1:250), goat polyclonal anti-platelet-derived growth factor receptor b (PDGFRb) antibody (AF1042: R&D Systems, Inc., 1:500), mouse monoclonal anti-microtubule associated protein (MAP2) antibody (M9942: Sigma-Aldrich Co. LLC, 1:250), and rabbit polyclonal anti-FLAG antibody (F7425: Sigma-Aldrich Co. LLC, 1:100).

Techniques: Microscopy, Staining, Marker, Derivative Assay

Journal: PloS one

Article Title: Effects of angiopoietin-1 on hemorrhagic transformation and cerebral edema after tissue plasminogen activator treatment for ischemic stroke in rats.

doi: 10.1371/journal.pone.0098639

Figure Lengend Snippet: Figure 4. COMP-Ang1 protein localization in the tPA-4h group visualized with a confocal laser microscope. From left to right: markers of cells that make up the blood brain barrier (a, e, i; red), neuronal cell (m; red), COMP-Ang1 protein (b, f, j, n; green), DAPI stain (c, g, k, o; blue), and a merged image (d, h, l, p). Immunostaining with an anti-FLAG antibody was performed on COMP-Ang1 proteins in order to differentiate them from endogenous Ang1. RECA1 is an endothelial cell marker protein; PDGFRb is a pericyte marker; GFAP is an astrocyte marker; and MAP2 is a neuronal cell marker. RECA1, rat endothelial cell antigen; PDGFRb, platelet-derived growth factor receptor; GFAP, glial fibrillary acidic protein; Ang1, angiopoietin-1. The scale bars are 10 mm. doi:10.1371/journal.pone.0098639.g004

Article Snippet: The primary antibodies that were used were a rabbit polyclonal anti-Ang1 antibody (ab93599: Abcam plc, 1:100), mouse monoclonal anti-RECA1 antibody (MCA-970R: AbD Serotec, 1:250), mouse monoclonal anti-glial fibrillary acidic protein (GFAP) antibody (3670: Cell Signaling Technology, Inc., 1:250), goat polyclonal anti-platelet-derived growth factor receptor b (PDGFRb) antibody (AF1042: R&D Systems, Inc., 1:500), mouse monoclonal anti-microtubule associated protein (MAP2) antibody (M9942: Sigma-Aldrich Co. LLC, 1:250), and rabbit polyclonal anti-FLAG antibody (F7425: Sigma-Aldrich Co. LLC, 1:100).

Techniques: Microscopy, Staining, Immunostaining, Marker, Derivative Assay