Log In Sign up
The largest database of trusted experimental protocols

Recombinant human c5a

Manufactured by R&D Systems
Visit Supplier
Sourced in United States

Recombinant human C5a is a protein produced using recombinant DNA technology. It is a component of the complement system, a part of the immune system that helps clear pathogens. The core function of recombinant human C5a is to serve as a research tool for studying the complement system and its role in various biological processes.

Automatically generated - may contain errors

Lab products found in correlation

Optilyse c solution, by Beckman Coulter (2 mentions) Facscanto 2 flow cytometer, by BD (2 mentions) Cytofix, by BD (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions) Simplyblue safestain, by Thermo Fisher Scientific (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Caco 2, by American Type Culture Collection (1 mentions) Wortmannin, by Merck Group (1 mentions) Tms diazomethane, by Merck Group (1 mentions) Byl719, by Active Motif (1 mentions) Murine tnf α, by R&D Systems (1 mentions) Murine gm csf, by Thermo Fisher Scientific (1 mentions) Fatty acid free bsa, by Merck Group (1 mentions) Hank s balanced salt solution, by Merck Group (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Luminol, by Merck Group (1 mentions) Cystine lactose electrolyte deficient, by Thermo Fisher Scientific (1 mentions) Cled agar, by Thermo Fisher Scientific (1 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (1 mentions)

8 protocols using recombinant human c5a

1

Neutrophil Functional Assay with C5a

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Purified neutrophils were resuspended to 1 × 107 viable cells/ml in IMDM, then diluted to 1 × 106/ml in IMDM containing 3% autologous serum and recombinant human C5a (R and D Systems, Abingdon) at 1–100 ng/ml or untreated control12 (link). Neutrophils were incubated by rotation (10 rpm) at 37 °C for 16 h prior to functional assays. Neutrophil viable counts were re-assessed by trypan blue exclusion and adjusted once again to 1 × 106 viable cells/ml.
Evans S.J., Roberts A.E., Morris A.C., Simpson A.J., Harris L.G., Mack D., Jenkins R.E, & Wilkinson T.S. (2020). Contrasting effects of linezolid on healthy and dysfunctional human neutrophils: reducing C5a-induced injury. Scientific Reports, 10, 16377.
+ Open protocol
+ Expand
2

Quantifying Bacterial C5a Peptidase Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
C5a peptidase activity was assessed by incubating bacterial strains with recombinant human C5a (R&D Systems, 2037-C5-025/CF). Stationary phase cultures of GBS and L. lactis were resuspended at 2 × 109 CFUs/mL and then incubated with 100 μg/mL of recombinant human C5a for 16 h at 37 °C. Following the incubation, NuPAGE LDS Sample Buffer 4× (Fisher, NP0007) was added to the sample supernatant and boiled for 5 min. Subsequently, the samples were run on a 4–12% Bis-Tris gel (Fisher) at 100 V and stained with SimplyBlue Safe Stain (Fisher, LC6065).
McCabe S., Bjånes E., Hendriks A., Wang Z., van Sorge N.M., Pill-Pepe L., Bautista L., Chu E., Codée J.D., Fairman J., Kapoor N., Uchiyama S, & Nizet V. (2023). The Group A Streptococcal Vaccine Candidate VAX-A1 Protects against Group B Streptococcus Infection via Cross-Reactive IgG Targeting Virulence Factor C5a Peptidase. Vaccines, 11(12), 1811.
+ Open protocol
+ Expand
3

Analyzing Transcriptional Changes in M-like Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
A human colon carcinoma cell line, Caco-2, and a human Burkitt's lymphoma cell line, Raji, were purchased from the American Type Culture Collection (Manassas, VA, USA). The in vitro human M-like cells which had been prepared as described in our previous study were treated with recombinant human C5a (R&D Systems, Minneapolis, MN, USA), lipopolysaccharide (LPS) and C5a, or Y. enterocolitica for 6 h and then total mRNA was purified using the RNeasy Plus mini kit (Qiagen, Hilden, Germany) according to the manufacturer's protocol (17 (link)). The transcript levels of each indicated genes were monitored by human cAMP/Ca2+ signaling pathway array kit (PAHS-066Z; Qiagen) according to the manufacturer's instructions using an Applied Biosystems 7500 Real-Time PCR Instrument System (Life Technology, Carlsbad, CA, USA).
Kim S.H, & Jang Y.S. (2017). Yersinia enterocolitica Exploits Signal Crosstalk between Complement 5a Receptor and Toll-like Receptor 1/2 and 4 to Avoid the Bacterial Clearance in M cells. Immune Network, 17(4), 228-236.
+ Open protocol
+ Expand
4

Murine Macrophage Signaling Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Bovine FGN, poly-RGD+, fMLP, luminol, TMS-diazomethane, wortmannin, fatty acid–free BSA, and HRP were from Sigma-Aldrich. Murine GM-CSF and M-CSF were from PeproTech, whereas murine TNF-α and recombinant human C5a were from R&D Systems. SRBCs in Alsever’s were from TCS Biosciences. Hanks’ balanced salt solution and Dulbecco’s phosphate-buffered saline (dPBS) with Ca2+ and Mg2+ were from Sigma. The isoform-selective PI3K inhibitors IC87114 and TGX221 were from Calbiochem, whereas BYL719 was from Active Biochem. All tissue culture products were from Invitrogen. All buffer components were from Sigma-Aldrich and were endotoxin-free or low-endotoxin, as available. Internal standards for lipid analysis, 1-heptadecanoyl-2-hexadecanoyl-sn-glycero-3-(phosphoinositol 3,4,5-trisphosphate) (C17:0/C16:0-PIP3, as a hepta sodium salt), and C17:0/C16:0-PI were synthesized at the Babraham Institute. All chemicals and solutions were of analytical reagent grade.
Houslay D.M., Anderson K.E., Chessa T., Kulkarni S., Fritsch R., Downward J., Backer J.M., Stephens L.R, & Hawkins P.T. (2016). Coincident signals from GPCRs and receptor tyrosine kinases are uniquely transduced by PI3Kβ in myeloid cells. Science signaling, 9(441), ra82.
+ Open protocol
+ Expand
5

Avdoralimab Inhibits C5a-induced Neutrophil Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Various concentrations of avdoralimab were added to the blood samples in culture-treated 96-well U-bottom plates, and incubated for 20 min at 37°C under an atmosphere containing 5% CO2. We then added 18 nM human recombinant C5a (R&D Systems) to the samples. Plates were incubated for 20 minutes at 37°C under an atmosphere containing 5% CO2. Samples were then stained for flow cytometry analysis with anti-CD16 FITC and anti-CD11b PE-Cy5 antibodies. Erythrocytes were lysed with Optilyse C solution (Beckman Coulter, A11895), according to the manufacturer’s protocol, and resuspended in CytoFix (BD Bioscience 554655) for fixation. Cells were then analyzed on a FACS Canto II flow cytometer (BD Biosciences) with FACS Diva software.
Carvelli J., Demaria O., Vély F., Batista L., Benmansour N.C., Fares J., Carpentier S., Thibult M.L., Morel A., Remark R., André P., Represa A., Piperoglou C., Cordier P.Y., Le Dault E., Guervilly C., Simeone P., Gainnier M., Morel Y., Ebbo M., Schleinitz N, & Vivier E. (2020). Association of COVID-19 inflammation with activation of the C5a-C5aR1 axis. Nature, 588(7836), 146-150.
+ Open protocol
+ Expand
6

Avdoralimab Inhibits C5a-induced Neutrophil Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Various concentrations of avdoralimab were added to the blood samples in culture-treated 96-well U-bottom plates, and incubated for 20 min at 37°C under an atmosphere containing 5% CO2. We then added 18 nM human recombinant C5a (R&D Systems) to the samples. Plates were incubated for 20 minutes at 37°C under an atmosphere containing 5% CO2. Samples were then stained for flow cytometry analysis with anti-CD16 FITC and anti-CD11b PE-Cy5 antibodies. Erythrocytes were lysed with Optilyse C solution (Beckman Coulter, A11895), according to the manufacturer’s protocol, and resuspended in CytoFix (BD Bioscience 554655) for fixation. Cells were then analyzed on a FACS Canto II flow cytometer (BD Biosciences) with FACS Diva software.
Carvelli J., Demaria O., Vély F., Batista L., Benmansour N.C., Fares J., Carpentier S., Thibult M.L., Morel A., Remark R., André P., Represa A., Piperoglou C., Cordier P.Y., Le Dault E., Guervilly C., Simeone P., Gainnier M., Morel Y., Ebbo M., Schleinitz N, & Vivier E. (2020). Association of COVID-19 inflammation with activation of the C5a-C5aR1 axis. Nature, 588(7836), 146-150.
+ Open protocol
+ Expand
7

Immunohistochemical analysis of C5aR1 expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following reagents were used in this study: tryptone, yeast extract, cystine lactose electrolyte deficient (CLED) agar (Oxoid, Basingstoke, UK); monoclonal rat anti-mouse Ly6G, polyclonal goat anti-mouse C5aR1 (M-19) (Santa Cruz Biotechnology, Heidelberg, Germany); polyclonal rabbit anti-mouse cytokeratin (Abcam); Alexa Fluor 488 donkey anti-goat IgG (Jackson Immunology Research Lab., West Grove, USA); fluorescein-labeled Galanthus nivalis lectin (GNL, which binds to mannosyl residues) (Vector Laboratories, Peterborough, UK); polyclonal rabbit anti-human ZO-1 and 4′,6-diamidino-2-phenylindole (DAPI) (Life Technologies Ltd., Beijing, China); protease inhibitor cocktail, tetramethylrhodamine (TRITC) (Sigma-Aldrich, St. Louis, USA); anti-phospho-ERK1/2 (Thr202/Tyr204), -IκB (Ser 32), and anti-ERK1/2, -IκB antibodies used for Western blot and signaling pathway studies (Cell Signaling Technology, Danvers, USA); cell culture medium, fetal calf serum, gentamicin, TRIzol, CountBright™ absolute counting beads, Fast SYBR® Green Master Mix, M-PER mammalian protein extraction reagent, RIPA lysis buffer and BCA protein assay kit (Thermo Fisher, Waltham, USA); human recombinant C5a (R&D Systems); C5aR1 peptide antagonist (PMX53, Ac-Phe-cyclo [Orn-Pro-dCha-Trp-Arg]); and control peptide (random sequence) (synthesized by GenScript, Shanghai, China).
Wu K.Y., Cao B., Wang C.X., Yang X.L., Zhao S.J., Diao T.Y., Lin L.R., Zhao G.X., Zhou W., Yang J.R, & Li K. (2022). The C5a/C5aR1 Axis Contributes to the Pathogenesis of Acute Cystitis Through Enhancement of Adhesion and Colonization of Uropathogenic E. coli. Frontiers in Cellular and Infection Microbiology, 12, 824505.
+ Open protocol
+ Expand
8

S1P Agonists, Antagonists, and Engineered Chaperone Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Specific S1P1 agonists, CYM-5442 (Santa Cruz Biotechnology) and SEW-2871 and a specific S1P1 antagonist, W146 (Cayman Chemical), were dissolved in DMSO . A novel engineered S1P chaperone, ApoM-Fc loaded with S1P- [19 ], was used in vitro (10 μg/ml) and in vivo (4 mg/kg IP). Human recombinant C5a (R&D Systems; 100 ng/ml) was used to activate PMNs. The following antibodies were used for western blot and/or immunofluorescence: ApoM (ab91656, Abcam), anti-VE-Cadherin (C19, Santa Cruz Biotechnology), and anti-p-MLC2 (T18/S19, Cell Signaling).
Burg N., Swendeman S., Worgall S., Hla T, & Salmon J.E. (2018). Sphingosine -1 Phosphate Receptor-1 signaling maintains endothelial cell barrier function and protects against immune complex-induced vascular injury. Arthritis & rheumatology (Hoboken, N.J.), 70(11), 1879-1889.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!