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recombinant mouse c3a  (R&D Systems)


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    R&D Systems recombinant mouse c3a
    Time-dependent <t>complement</t> <t>activation</t> and anaphylatoxin receptor expression in ALI cultures exposed to HDM. ( a ) C3 and <t>C3a</t> concentrations in cell lysates and the SN of unstimulated (steady-state) ALI cultures and 24, 48 and 72 h after HDM exposure (steady-state, 24, 48 and 72 h: n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001. ( b ) HDM and HI-HDM mediated cleavage of hC3 into hC3a ( n = 3 independent experiments). Data are shown as mean ± SEM. Data were analyzed using an unpaired T -test. *** p < 0.001. ( c ) C5 and C5a concentrations in cell lysates and the SN of steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). ( d ) Comparison of C3 and C5 ( left panel ), C3a and C5a ( right panel ) concentrations in cell lysates from steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). Data were analyzed using two-way ANOVA with Šidák’s multiple comparisons test. **** p < 0.0001. ( e ) Immunofluorescence analysis of C3aR and C5aR1 expression in unstimulated ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM ( n = 3). Data were analyzed using one-way ANOVA with Holm–Šidák’s multiple comparisons test. * p < 0.05; ** p < 0.01. The number of experiments shown in ( a , c – e ) refers to biological replicates derived from 6 independent preparations of trachea, each representing a separate cell isolation and culture.
    Recombinant Mouse C3a, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+mouse+c3a/Recombinant+Mouse+Complement+Component+C3a+Protein%2C+CF/pmc12563611-130-14-17
    Average 93 stars, based on 15 article reviews
    recombinant mouse c3a - by Bioz Stars, 2026-09
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    Images

    1) Product Images from "House Dust Mite Nebulization Drives Alarmin and Complement Activation in a Murine Tracheal Air–Liquid Interface Culture System"

    Article Title: House Dust Mite Nebulization Drives Alarmin and Complement Activation in a Murine Tracheal Air–Liquid Interface Culture System

    Journal: Cells

    doi: 10.3390/cells14201598

    Time-dependent complement activation and anaphylatoxin receptor expression in ALI cultures exposed to HDM. ( a ) C3 and C3a concentrations in cell lysates and the SN of unstimulated (steady-state) ALI cultures and 24, 48 and 72 h after HDM exposure (steady-state, 24, 48 and 72 h: n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001. ( b ) HDM and HI-HDM mediated cleavage of hC3 into hC3a ( n = 3 independent experiments). Data are shown as mean ± SEM. Data were analyzed using an unpaired T -test. *** p < 0.001. ( c ) C5 and C5a concentrations in cell lysates and the SN of steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). ( d ) Comparison of C3 and C5 ( left panel ), C3a and C5a ( right panel ) concentrations in cell lysates from steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). Data were analyzed using two-way ANOVA with Šidák’s multiple comparisons test. **** p < 0.0001. ( e ) Immunofluorescence analysis of C3aR and C5aR1 expression in unstimulated ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM ( n = 3). Data were analyzed using one-way ANOVA with Holm–Šidák’s multiple comparisons test. * p < 0.05; ** p < 0.01. The number of experiments shown in ( a , c – e ) refers to biological replicates derived from 6 independent preparations of trachea, each representing a separate cell isolation and culture.
    Figure Legend Snippet: Time-dependent complement activation and anaphylatoxin receptor expression in ALI cultures exposed to HDM. ( a ) C3 and C3a concentrations in cell lysates and the SN of unstimulated (steady-state) ALI cultures and 24, 48 and 72 h after HDM exposure (steady-state, 24, 48 and 72 h: n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001. ( b ) HDM and HI-HDM mediated cleavage of hC3 into hC3a ( n = 3 independent experiments). Data are shown as mean ± SEM. Data were analyzed using an unpaired T -test. *** p < 0.001. ( c ) C5 and C5a concentrations in cell lysates and the SN of steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). ( d ) Comparison of C3 and C5 ( left panel ), C3a and C5a ( right panel ) concentrations in cell lysates from steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). Data were analyzed using two-way ANOVA with Šidák’s multiple comparisons test. **** p < 0.0001. ( e ) Immunofluorescence analysis of C3aR and C5aR1 expression in unstimulated ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM ( n = 3). Data were analyzed using one-way ANOVA with Holm–Šidák’s multiple comparisons test. * p < 0.05; ** p < 0.01. The number of experiments shown in ( a , c – e ) refers to biological replicates derived from 6 independent preparations of trachea, each representing a separate cell isolation and culture.

    Techniques Used: Activation Assay, Expressing, Comparison, Immunofluorescence, Derivative Assay, Cell Isolation

    Related Articles

    Recombinant:

    Article Title: Complement C3a activates astrocytes to promote medulloblastoma progression through TNF-α.
    Article Snippet: Recombinant mouse C3a, TNF-α and IL-6 were purchased from Novoprotein. .. In addition, recombinant mouse C3a from R&D Systems was used in some of the experiments as indicated. ..

    Article Title: Macrophage to myofibroblast transition contributes to subretinal fibrosis secondary to neovascular age-related macular degeneration.
    Article Snippet: Seven days later, the phenotype of BMDMs was checked by flow cytometry (FACS BD CANTO II, BD Biosciences, San Jose, USA) using the following antibodies: F4/80 conjugated eFluor 450 (Thermo Fisher Scientific, Cat no: 48-4801-80) and CD11b conjugated PE-Cy7 (BD Biosciences, Cat no: 552850). .. BMDMs cultured in 10% FCS were treated with 10 ng/ ml recombinant mouse C3a (Cat No: 8085-C3) or C5a (Cat No: 2150-C5-025/CF) for 48–96 h. Ten nanograms per milliliter of TGF-β1 (Cat No: 7666-MB-005) (all from R&D Systems, Abingdon, UK) was used as a positive control. .. C3aR antagonist (C3aRA) SB 290157 (trifluoroacetate salt) (Cayman Chemical, Michigan, USA, Cat No: 15783) was dissolved in DMSO according to the manufacturer’s instructions.

    Article Title: Complement C3a activates astrocytes to promote medulloblastoma progression through TNF-α
    Article Snippet: Recombinant mouse C3a, TNF-α and IL-6 were purchased from Novoprotein. .. In addition, recombinant mouse C3a from R&D Systems was used in some of the experiments as indicated. ..

    Article Title: Genome-scale CRISPR screening reveals that C3aR signaling is critical for rapid capture of fungi by macrophages.
    Article Snippet: .. BMDMs were pre-incubated in serum-free media with varying concentrations of recombinant mouse C3a (R&D systems) or with 20% FBS for 1 h, then infected with A.Hc or B. Zymosan at an MOI of 5 for 30 min. Phagocytosis was assessed by flow cytometry (n = 2 biological replicates). ..

    Article Title: Endothelial progenitor cell transplantation alleviated ischemic brain injury via inhibiting C3/C3aR pathway in mice
    Article Snippet: Microglia culture and C3a treatment BV2, immortalized mouse microglial cells, were plated in the 12-well (Corning Incorporated) with a density of 7 × 10 4 per well and cultured in DMEM with 5% heat-inactivated FBS. .. When the density of microglia reached about 50% confluence (about one day), recombinant mouse C3a (R&D) with a final concentration of 100 Mm was added to the culture medium. ..

    Article Title: TLQP-21 is a low potency partial C3aR activator on human primary macrophages
    Article Snippet: The C3aR inhibitor, SB290157 trifluoroacetate salt, and purified human C3a were purchased from Merck (Perth, Australia). .. Recombinant mouse C3a was purchased from R&D Systems (Minneapolis, USA). .. Bovine serum albumin (BSA) was purchased from Merck (Perth, Australia).

    Article Title: House Dust Mite Nebulization Drives Alarmin and Complement Activation in a Murine Tracheal Air–Liquid Interface Culture System
    Article Snippet: .. Plates were washed four times with 0.05% Tween 20 in PBS; serial dilutions of recombinant mouse C3a (R&D Systems #8085-C3-025—Minneapolis, MN, USA) were performed; and samples were added (25 μL/well) and incubated at RT for 90 min. After washing four times with 0.05% Tween 20 in PBS, 25 μL/well of 1 μg/mL in 1% BSA/PBS biotinylated anti-mouse C3a detection antibody (clone I87-419, BD Biosciences #558251—San Jose, CA, USA) was added and incubated for 1 h at RT. ..

    Cell Culture:

    Article Title: Macrophage to myofibroblast transition contributes to subretinal fibrosis secondary to neovascular age-related macular degeneration.
    Article Snippet: Seven days later, the phenotype of BMDMs was checked by flow cytometry (FACS BD CANTO II, BD Biosciences, San Jose, USA) using the following antibodies: F4/80 conjugated eFluor 450 (Thermo Fisher Scientific, Cat no: 48-4801-80) and CD11b conjugated PE-Cy7 (BD Biosciences, Cat no: 552850). .. BMDMs cultured in 10% FCS were treated with 10 ng/ ml recombinant mouse C3a (Cat No: 8085-C3) or C5a (Cat No: 2150-C5-025/CF) for 48–96 h. Ten nanograms per milliliter of TGF-β1 (Cat No: 7666-MB-005) (all from R&D Systems, Abingdon, UK) was used as a positive control. .. C3aR antagonist (C3aRA) SB 290157 (trifluoroacetate salt) (Cayman Chemical, Michigan, USA, Cat No: 15783) was dissolved in DMSO according to the manufacturer’s instructions.

    Positive Control:

    Article Title: Macrophage to myofibroblast transition contributes to subretinal fibrosis secondary to neovascular age-related macular degeneration.
    Article Snippet: Seven days later, the phenotype of BMDMs was checked by flow cytometry (FACS BD CANTO II, BD Biosciences, San Jose, USA) using the following antibodies: F4/80 conjugated eFluor 450 (Thermo Fisher Scientific, Cat no: 48-4801-80) and CD11b conjugated PE-Cy7 (BD Biosciences, Cat no: 552850). .. BMDMs cultured in 10% FCS were treated with 10 ng/ ml recombinant mouse C3a (Cat No: 8085-C3) or C5a (Cat No: 2150-C5-025/CF) for 48–96 h. Ten nanograms per milliliter of TGF-β1 (Cat No: 7666-MB-005) (all from R&D Systems, Abingdon, UK) was used as a positive control. .. C3aR antagonist (C3aRA) SB 290157 (trifluoroacetate salt) (Cayman Chemical, Michigan, USA, Cat No: 15783) was dissolved in DMSO according to the manufacturer’s instructions.

    Infection:

    Article Title: Genome-scale CRISPR screening reveals that C3aR signaling is critical for rapid capture of fungi by macrophages.
    Article Snippet: .. BMDMs were pre-incubated in serum-free media with varying concentrations of recombinant mouse C3a (R&D systems) or with 20% FBS for 1 h, then infected with A.Hc or B. Zymosan at an MOI of 5 for 30 min. Phagocytosis was assessed by flow cytometry (n = 2 biological replicates). ..

    Flow Cytometry:

    Article Title: Genome-scale CRISPR screening reveals that C3aR signaling is critical for rapid capture of fungi by macrophages.
    Article Snippet: .. BMDMs were pre-incubated in serum-free media with varying concentrations of recombinant mouse C3a (R&D systems) or with 20% FBS for 1 h, then infected with A.Hc or B. Zymosan at an MOI of 5 for 30 min. Phagocytosis was assessed by flow cytometry (n = 2 biological replicates). ..

    Concentration Assay:

    Article Title: Endothelial progenitor cell transplantation alleviated ischemic brain injury via inhibiting C3/C3aR pathway in mice
    Article Snippet: Microglia culture and C3a treatment BV2, immortalized mouse microglial cells, were plated in the 12-well (Corning Incorporated) with a density of 7 × 10 4 per well and cultured in DMEM with 5% heat-inactivated FBS. .. When the density of microglia reached about 50% confluence (about one day), recombinant mouse C3a (R&D) with a final concentration of 100 Mm was added to the culture medium. ..

    Incubation:

    Article Title: House Dust Mite Nebulization Drives Alarmin and Complement Activation in a Murine Tracheal Air–Liquid Interface Culture System
    Article Snippet: .. Plates were washed four times with 0.05% Tween 20 in PBS; serial dilutions of recombinant mouse C3a (R&D Systems #8085-C3-025—Minneapolis, MN, USA) were performed; and samples were added (25 μL/well) and incubated at RT for 90 min. After washing four times with 0.05% Tween 20 in PBS, 25 μL/well of 1 μg/mL in 1% BSA/PBS biotinylated anti-mouse C3a detection antibody (clone I87-419, BD Biosciences #558251—San Jose, CA, USA) was added and incubated for 1 h at RT. ..



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    Time-dependent complement activation and anaphylatoxin receptor expression in ALI cultures exposed to HDM. ( a ) C3 and C3a concentrations in cell lysates and the SN of unstimulated (steady-state) ALI cultures and 24, 48 and 72 h after HDM exposure (steady-state, 24, 48 and 72 h: n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001. ( b ) HDM and HI-HDM mediated cleavage of hC3 into hC3a ( n = 3 independent experiments). Data are shown as mean ± SEM. Data were analyzed using an unpaired T -test. *** p < 0.001. ( c ) C5 and C5a concentrations in cell lysates and the SN of steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). ( d ) Comparison of C3 and C5 ( left panel ), C3a and C5a ( right panel ) concentrations in cell lysates from steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). Data were analyzed using two-way ANOVA with Šidák’s multiple comparisons test. **** p < 0.0001. ( e ) Immunofluorescence analysis of C3aR and C5aR1 expression in unstimulated ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM ( n = 3). Data were analyzed using one-way ANOVA with Holm–Šidák’s multiple comparisons test. * p < 0.05; ** p < 0.01. The number of experiments shown in ( a , c – e ) refers to biological replicates derived from 6 independent preparations of trachea, each representing a separate cell isolation and culture.

    Journal: Cells

    Article Title: House Dust Mite Nebulization Drives Alarmin and Complement Activation in a Murine Tracheal Air–Liquid Interface Culture System

    doi: 10.3390/cells14201598

    Figure Lengend Snippet: Time-dependent complement activation and anaphylatoxin receptor expression in ALI cultures exposed to HDM. ( a ) C3 and C3a concentrations in cell lysates and the SN of unstimulated (steady-state) ALI cultures and 24, 48 and 72 h after HDM exposure (steady-state, 24, 48 and 72 h: n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001. ( b ) HDM and HI-HDM mediated cleavage of hC3 into hC3a ( n = 3 independent experiments). Data are shown as mean ± SEM. Data were analyzed using an unpaired T -test. *** p < 0.001. ( c ) C5 and C5a concentrations in cell lysates and the SN of steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). ( d ) Comparison of C3 and C5 ( left panel ), C3a and C5a ( right panel ) concentrations in cell lysates from steady-state ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM (steady-state, 24, 48 and 72 h: n = 6). Data were analyzed using two-way ANOVA with Šidák’s multiple comparisons test. **** p < 0.0001. ( e ) Immunofluorescence analysis of C3aR and C5aR1 expression in unstimulated ALI cultures and 24, 48 and 72 h after HDM exposure. Data are shown as mean ± SEM ( n = 3). Data were analyzed using one-way ANOVA with Holm–Šidák’s multiple comparisons test. * p < 0.05; ** p < 0.01. The number of experiments shown in ( a , c – e ) refers to biological replicates derived from 6 independent preparations of trachea, each representing a separate cell isolation and culture.

    Article Snippet: Plates were washed four times with 0.05% Tween 20 in PBS; serial dilutions of recombinant mouse C3a (R&D Systems #8085-C3-025—Minneapolis, MN, USA) were performed; and samples were added (25 μL/well) and incubated at RT for 90 min. After washing four times with 0.05% Tween 20 in PBS, 25 μL/well of 1 μg/mL in 1% BSA/PBS biotinylated anti-mouse C3a detection antibody (clone I87-419, BD Biosciences #558251—San Jose, CA, USA) was added and incubated for 1 h at RT.

    Techniques: Activation Assay, Expressing, Comparison, Immunofluorescence, Derivative Assay, Cell Isolation

    Microglial C3a – C3aR signaling promotes glycolysis and phagocytosis. Young microglia (postnatal day 0–2) were treated with 10 nM recombinant mouse C3a. ( a - d ) p-AKT (Ser473), p-mTOR (Ser2448) and HIF1α were assessed by Western blotting at the indicated timepoints (0–24 h), and expression was normalized to total AKT, total mTOR and β-actin, respectively. n = 5 replicates. Data are presented as mean (SEM). * p < 0.05, ** p < 0.01, *** p < 0.001 (one-way ANOVA). ( e - g ) Seahorse assays were used to evaluate real-time glycolytic rate in young microglia (postnatal day 0–2, pooled male and female) after 18 h in vitro treatment with 10 nM recombinant mouse C3a. Stimuli were added as indicated ( e ), and basal glycolysis ( f ) and compensatory glycolysis ( g ) were determined by calculating the glycolytic Proton Efflux Rate (e; glycoPER). n = 5/group. # p < 0.05 (unpaired t-test). ( h , i ) Flow cytometry analysis was performed to assess phagocytosis of FITC-fAβ 1−42 . Rapamycin (50 µM) or 2-DG (5 mM) were pre- (1 h) and co-treated (18 h) with C3a. Proportion of phagocytic cells ( h ) was assessed by evaluating FITC-positive microglia, and FITC MFI was evaluated in total live microglia ( i ). Data are presented as mean (SEM). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (one-way ANOVA)

    Journal: Journal of Neuroinflammation

    Article Title: Microglia undergo sex-dimorphic transcriptional and metabolic rewiring during aging

    doi: 10.1186/s12974-024-03130-7

    Figure Lengend Snippet: Microglial C3a – C3aR signaling promotes glycolysis and phagocytosis. Young microglia (postnatal day 0–2) were treated with 10 nM recombinant mouse C3a. ( a - d ) p-AKT (Ser473), p-mTOR (Ser2448) and HIF1α were assessed by Western blotting at the indicated timepoints (0–24 h), and expression was normalized to total AKT, total mTOR and β-actin, respectively. n = 5 replicates. Data are presented as mean (SEM). * p < 0.05, ** p < 0.01, *** p < 0.001 (one-way ANOVA). ( e - g ) Seahorse assays were used to evaluate real-time glycolytic rate in young microglia (postnatal day 0–2, pooled male and female) after 18 h in vitro treatment with 10 nM recombinant mouse C3a. Stimuli were added as indicated ( e ), and basal glycolysis ( f ) and compensatory glycolysis ( g ) were determined by calculating the glycolytic Proton Efflux Rate (e; glycoPER). n = 5/group. # p < 0.05 (unpaired t-test). ( h , i ) Flow cytometry analysis was performed to assess phagocytosis of FITC-fAβ 1−42 . Rapamycin (50 µM) or 2-DG (5 mM) were pre- (1 h) and co-treated (18 h) with C3a. Proportion of phagocytic cells ( h ) was assessed by evaluating FITC-positive microglia, and FITC MFI was evaluated in total live microglia ( i ). Data are presented as mean (SEM). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (one-way ANOVA)

    Article Snippet: Primary microglia were stimulated with vehicle (PBS) or recombinant mouse C3a (10 nM; Peprotech) for the indicated time as in each figure legend.

    Techniques: Recombinant, Western Blot, Expressing, In Vitro, Flow Cytometry