Invitrogen

Complement C3 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

8 Published Figures

15 References

View all (18) Complement C3 antibodies

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Protocols

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Complement C3 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Complement C3 was performed using 70% confluent log phase Hep G2 cells treated with 10 ng/mL of LPS for 18 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Complement C3 Polyclonal Antibody (Product # PA5-21349) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents untreated cells with weak cytoplasmic signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Complement C3 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — C3 antibody [C3], C-term detects C3 protein at cytoplasm in mouse brain by immunohistochemical analysis. Sample: Paraffin-embedded mouse brain. C3 antibody [C3], C-term (Product # PA5-21349) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Complement C3 Antibody in Western Blot (WB) — Western blot was performed using Anti-Complement C3 Polyclonal Antibody (Product # PA5-21349) and a 187 kDa band corresponding to Complement C3 was observed across cell lines tested. The expression of Complement C3 was upregulated with LPS and IL-1beta treatment. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with Protein transport inhibitor (1X for 4hr) (Lane 2), Hep G2 treated with LPS (10 ng/mL for 18hr) and Protein transport inhibitor (1X for 4 hr) (Lane 3), A549 (Lane 4) and A549 treated with IL-1beta (5 ng/mL 48hr) (Lane 5) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Complement C3 Antibody in Immunocytochemistry, Immunohistochemistry (ICC/IF, IHC) — Decorin+ astrocytes are associated with vascular amyloid in vivo. a Triple immunofluorescence of amyloid (Thio-S, green), astrocytes (GFAP, cyan), and Decorin (red) in the cortex of 15-month-old Tg-FDD, APP/PS1 mice and WT controls. Major presence of Decorin immunoreactivity is observed in the perivascular region of Thio-S positive vasculature in Tg-FDD mice. Yellow arrows indicate Decorin + , GFAP + cells and purple arrows indicate Decorin-, GFAP + cells. Colocalization analysis (CC) was performed to determine pixel intensity correlation between GFAP and Decorin. White pixels indicate colocalization between GFAP and Decorin signal and white dashed arrows represent distance in plot profiles showing overlapping intensities of astrocytes (GFAP, cyan) and Decorin (red). b Quantification of the proportion of Decorin + cells in Tg-FDD, APP/PS1 and WT mice. c Triple immunofluorescence of astrocytes (GFAP, green), complement component 3 (C3, cyan), and Decorin (red) in the cortex of 15-month-old Tg-FDD and APP/PS1 mice. Yellow arrows indicate C3 + , Decorin + , GFAP + cells and purple arrows indicate C3 + , Decorin-, GFAP + cells. d Quantification of the proportion of C3 + Decorin + astrocytes versus C3 + astrocytes in Tg-FDD, APP/PS1 and WT mice. For quantifications, eight to ten images were used for each experiment ( n = 3), and 200 cells were counted. All are representative images of 15-month-old Tg-FDD, APP/PS1 or WT mice. Scale bar 25 or 50 mum, respectively.

Complement C3 Antibody in Immunohistochemistry (IHC) — Fig. 3 A1 Astrocytes are highly abundant in Tg-FDD. a Double-stained immunofluorescent images of complement component 3 (C3, red) and astrocytes (GFAP, green) in the hippocampus of WT and Tg-FDD mice. C3 and GFAP immunoreactivity overlay (Merge). b Quantification of C3 + GFAP + cell % from 3 regions per animal of the hippocampus of 4 WT and 3 Tg-FDD mice. c Quantification of % colocalization area from the hippocampus of 4 WT and 3 Tg-FDD mice. d Double-stained immunofluorescent images of complement component 3 (C3, red) and astrocytes (GFAP, green) in the cerebellum of WT and Tg-FDD mice. C3 and GFAP immunoreactivity overlay (Merge). e Quantification of C3 + GFAP + cells % from 3 regions per animal of the cerebellum of 4 WT and 3 Tg-FDD mice. f Quantification of % colocalization area from the cerebellum of 4 WT and 3 Tg-FDD mice. a and d are representative images of the brain regions of 9-month-old mice. Colocalization analysis (CC) was performed to determine pixel intensity correlation between C3 and GFAP signals. White pixels indicate colocalization between C3 and GFAP signal. Plot profiles of representative C3 and GFAP intensities show higher overlapping of C3 with GFAP in Tg-FDD mice in comparison with WT mice, indicating an increase of C3 positive astrocytes. Results are shown as the mean +- SEM of n = 3-4. Asterisks indicate significant differences, where **** p < 0.0001 by unpaired Student's t test. Scale bar 100 mum

Complement C3 Antibody in Flow Cytometry (Flow) — FIG 9 APRc protects E. coli from complement-mediated killing, and interaction with IgG contributes to serum resistance. (A) Western blot analysis with anti-APRc antibody of total protein extracts from BL21 Star(DE3) E. coli strain expressing the empty vector backbone pET28a (pET) or the plasmid encoding untagged full-length APRc wild type (APRcFL) and the corresponding catalytic mutant (APRcFL_D140N). (B) The serum-sensitive BL21 Star(DE3) E. coli strain expressing the empty vector (pET) or the plasmid encoding APRcFL and APRcFL_D140N was independently incubated 1:1 with PBS or in PBS containing 40% NHS for 1 h. After incubation, the samples were serially diluted, plated onto LB agar plates, and incubated overnight at 37degC. The average number of CFU per milliliter was calculated from the replicate plate counts. The data are presented as survival rate, which was calculated as the percentage of the original cell number at T 0 (considered 100% survival). Data represent the mean +- SD from 4 independent biological replicates. Significance was determined using a one-way ANOVA followed by Tukey multiple-comparison test using GraphPad Prism 8 (***, P < 0.001; ****, P < 0.0001). (C) BL21 Star(DE3) E. coli strain expressing the empty vector (pET) or the plasmid encoding APRcFL_D140N was independently incubated 1:1 with PBS or in PBS containing 40% NHS or 40% NHSDeltaIgG/IgM for 1 h. After incubation, samples were treated as described for panel B. Data represent the mean +- SD from 4

Complement C3 Antibody — Detection of altered subcellular localization of the target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis using Complement C3 Polyclonal Antibody (Product # PA5-21349), shows increased expression of protein in Hep G2 cell line upon treatment with LPS. {TM}

Product Details

PA5-21349

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	View 5 publications 5 publications
Immunohistochemistry (IHC)	-	View 4 publications 4 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	View 4 publications 4 publications
Flow Cytometry (Flow)	-	View 2 publications 2 publications
Immunoprecipitation (IP)	1:100-1:500	-

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein encompassing a sequence within the C-terminal region of human C3 (aa 1448-1655) View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.08 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol, 1% BSA
Contains	0.025% ProClin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_11153785

Product Specific Information

Recommended positive controls: Huh7, HepG2.

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

C3 is a key component of the complement system since classical and alternative activation pathways merge at the C3 activation step when C3 is split into C3a and C3b. The molecular mass of C3 is 185 kDa and it consists of two chains (110 kDa and 75 kDa) held together by disulfide bonds.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Activated endothelial cells induce a distinct type of astrocytic reactivity.

Communications biology

Taylor X,Cisternas P,Jury N,Martinez P,Huang X,You Y,Redding-Ochoa J,Vidal R,Zhang J,Troncoso J,Lasagna-Reeves CA

Published figure using Complement C3 polyclonal antibody (Product # PA5-21349) in Immunocytochemistry

Tue Mar 29 00:00:00 EDT 2022

Activated endothelial cells induce a distinct type of astrocytic reactivity.

Communications biology

Taylor X,Cisternas P,Jury N,Martinez P,Huang X,You Y,Redding-Ochoa J,Vidal R,Zhang J,Troncoso J,Lasagna-Reeves CA

Published figure using Complement C3 polyclonal antibody (Product # PA5-21349) in Immunocytochemistry

Tue Mar 29 00:00:00 EDT 2022

The Retropepsin-Type Protease APRc as a Novel Ig-Binding Protein and Moonlighting Immune Evasion Factor of Rickettsia.

mBio

Curto P,Barro A,Almeida C,Vieira-Pires RS,Simões I

Published figure using Complement C3 polyclonal antibody (Product # PA5-21349) in Flow Cytometry

Tue Dec 21 00:00:00 EST 2021

Bacillus anthracis Poly-γ-D-Glutamate Capsule Inhibits Opsonic Phagocytosis by Impeding Complement Activation.

Frontiers in immunology

Sharma S,Bhatnagar R,Gaur D

PA5-21349 was used in Western Blotting to demonstrate the mechanism through which the B. anthracis capsule imparts a survival advantage to the active bacteria.

Thu Mar 04 00:00:00 EST 2021

Ceria nanoparticles ameliorate white matter injury after intracerebral hemorrhage: microglia-astrocyte involvement in remyelination.

Journal of neuroinflammation

Zheng J,Lu J,Mei S,Wu H,Sun Z,Fang Y,Xu S,Wang X,Shi L,Xu W,Chen S,Yu J,Liang F,Zhang J

PA5-21349 was used in Immunohistochemistry to reveal a new mechanism in white matter injury after ICH: ICH induces M1 microglia and A1 astrocyte through ROS-induced NF-κB p65 translocation that hinders OPC maturation.

Mon Feb 15 00:00:00 EST 2021

Ceria nanoparticles ameliorate white matter injury after intracerebral hemorrhage: microglia-astrocyte involvement in remyelination.

Journal of neuroinflammation

Zheng J,Lu J,Mei S,Wu H,Sun Z,Fang Y,Xu S,Wang X,Shi L,Xu W,Chen S,Yu J,Liang F,Zhang J

Mon Feb 15 00:00:00 EST 2021

Complement in Reproductive White Adipose Tissue Characterizes the Obese Preeclamptic-Like BPH/5 Mouse Prior to and During Pregnancy.

Biology

Olson KN,Reijnders D,Gomes VCL,Hebert RC,Liu CC,Stephens JM,Redman LM,Douglas NC,Sones JL

PA5-21349 was used in Western Blotting to explore potential contribution of inflammatory mediators in visceral white adipose tissue (WAT) to Preeclampsia.

Tue Sep 22 00:00:00 EDT 2020

Complement in Reproductive White Adipose Tissue Characterizes the Obese Preeclamptic-Like BPH/5 Mouse Prior to and During Pregnancy.

Biology

Olson KN,Reijnders D,Gomes VCL,Hebert RC,Liu CC,Stephens JM,Redman LM,Douglas NC,Sones JL

PA5-21349 was used in Western Blotting to explore potential contribution of inflammatory mediators in visceral white adipose tissue (WAT) to Preeclampsia.

Tue Sep 22 00:00:00 EDT 2020

Mesenchymal stem cell-derived exosomes as a nanotherapeutic agent for amelioration of inflammation-induced astrocyte alterations in mice.

Theranostics

Xian P,Hei Y,Wang R,Wang T,Yang J,Li J,Di Z,Liu Z,Baskys A,Liu W,Wu S,Long Q

PA5-21349 was used in Immunohistochemistry to show that MSC-Exo ameliorate inflammation-induced astrocyte alterations and that the Nrf2-NF-κB signaling pathway is involved in regulating astrocyte activation in mice.

Mon Aug 31 00:00:00 EDT 2020

Mesenchymal stem cell-derived exosomes as a nanotherapeutic agent for amelioration of inflammation-induced astrocyte alterations in mice.

Theranostics

Xian P,Hei Y,Wang R,Wang T,Yang J,Li J,Di Z,Liu Z,Baskys A,Liu W,Wu S,Long Q

Mon Aug 31 00:00:00 EDT 2020

Mesenchymal stem cell-derived exosomes as a nanotherapeutic agent for amelioration of inflammation-induced astrocyte alterations in mice.

Theranostics

Xian P,Hei Y,Wang R,Wang T,Yang J,Li J,Di Z,Liu Z,Baskys A,Liu W,Wu S,Long Q

Mon Aug 31 00:00:00 EDT 2020

A1 reactive astrocytes and a loss of TREM2 are associated with an early stage of pathology in a mouse model of cerebral amyloid angiopathy.

Journal of neuroinflammation

Taylor X,Cisternas P,You Y,You Y,Xiang S,Marambio Y,Zhang J,Vidal R,Lasagna-Reeves CA

PA5-21349 was used in Immunocytochemistry-immunoflourescence to demonstrate that initial glial response associated with early-stage CAA is characterized by the upregulation of A1 astrocytes without significant microglial reactivity.

Sat Jul 25 00:00:00 EDT 2020

A1 reactive astrocytes and a loss of TREM2 are associated with an early stage of pathology in a mouse model of cerebral amyloid angiopathy.

Journal of neuroinflammation

Taylor X,Cisternas P,You Y,You Y,Xiang S,Marambio Y,Zhang J,Vidal R,Lasagna-Reeves CA

Sat Jul 25 00:00:00 EDT 2020

Loss of Complement Factor H impairs antioxidant capacity and energy metabolism of human RPE cells.

Scientific reports

Armento A,Honisch S,Panagiotakopoulou V,Sonntag I,Jacob A,Bolz S,Kilger E,Deleidi M,Clark S,Ueffing M

PA5-21349 was used in Western Blotting to suggest that endogenously produced FH contributes to transcriptional and metabolic homeostasis and protects RPE cells from oxidative stress, highlighting a novel role of FH in AMD pathogenesis.

Thu Jun 25 00:00:00 EDT 2020

IgE Contributes to Atherosclerosis and Obesity by Affecting Macrophage Polarization, Macrophage Protein Network, and Foam Cell Formation.

Arteriosclerosis, thrombosis, and vascular biology

Zhang X,Li J,Luo S,Wang M,Huang Q,Deng Z,de Febbo C,Daoui A,Liew PX,Sukhova GK,Metso J,Jauhiainen M,Shi GP,Guo J

PA5-21349 was used in Western Blotting to demonstrate that IgE deficiency reduced atherosclerosis lesion burden, lesion lipid deposition, smooth muscle cell and endothelial cell contents, chemokine MCP (monocyte chemoattractant protein)-1 expression and macrophage accumulation.

Sun Mar 01 00:00:00 EST 2020

Immunity against the Obligate Intracellular Bacterial Pathogen Rickettsia australis Requires a Functional Complement System.

Infection and immunity

Riley SP,Fish AI,Del Piero F,Martinez JJ

PA5-21349 was used in Flow cytometry/Cell sorting to look into when the complement system is activated during infection with Rickettsia australis in mice and how immunity can be drawn from this.

Fri Jun 01 00:00:00 EDT 2018

Bioinformatics

Protein Aliases: Acylation stimulating protein; acylation-stimulating protein cleavage product; ASP; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1; C3-beta-c; C3a anaphylatoxin; C3adesArg; C3bc; Complement C3; Complement C3 alpha chain; Complement C3 beta chain; Complement C3b alpha' chain; Complement C3c alpha' chain fragment 1; Complement C3c alpha' chain fragment 2; Complement C3d fragment; Complement C3dg fragment; Complement C3f fragment; Complement C3g fragment; complement component C3a; complement component C3b; epididymis secretory sperm binding protein Li 62p; HSE-MSF; prepro-C3

Gene Aliases: AHUS5; AI255234; ARMD9; ASP; C3; C3a; C3b; CPAMD1; HEL-S-62p; HSE-MSF; Plp

UniProt ID: (Human) P01024, (Mouse) P01027

Entrez Gene ID: (Human) 718, (Mouse) 12266

Molecular Function: protease inhibitor