Human Complement Component C5a DuoSet ELISA

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Human Complement Component C5a ELISA Standard Curve
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Product Details
Citations (22)
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Human Complement Component C5a DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Complement ComponentC5a. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.


Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human Complement Component C5a ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Complement Component C5a

Human complement 5a (C5a) is an enzymatically generated glycoprotein that belongs to a family of structurally and functionally related proteins known as anaphylatoxins. C5a is a 74 amino acid (aa) peptide that is created by the activity of C5a convertase on the C5 alpha-chain. Human C5a has four alpha-helices plus three intrachain disulfide bonds that create a triple loop structure.

Entrez Gene IDs:
727 (Human); 15139 (Mouse)
Alternate Names:
C5; C5A; Complement Component C5a; HE

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.


Citations for Human Complement Component C5a DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

22 Citations: Showing 1 - 10
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  1. Evaluation of Patients with Vaccine Allergies Prior to mRNA-Based COVID-19 Vaccination
    Authors: XR Lim, JWL Tan, GYL Chan, J Hou, L Xie, VHL Goh, J Boon, SSM Lee, CM Teo, SC Tan, KP Leong, BYH Thong, BPL Leung
    Vaccines, 2022;10(7):.
    Species: Human
    Sample Types: Serum
  2. Soluble MAC is primarily released from MAC-resistant bacteria that potently convert complement component C5
    Authors: DJ Doorduijn, MV Lukassen, MFL van 't Wou, V Franc, M Ruyken, BW Bardoel, AJR Heck, SHM Rooijakker
    Elife, 2022;11(0):.
    Species: Human
    Sample Types: Serum
  3. Intracellular complement C5a/C5aR1 stabilizes beta-catenin to promote colorectal tumorigenesis
    Authors: P Ding, Y Xu, L Li, X Lv, L Li, J Chen, D Zhou, X Wang, Q Wang, W Zhang, T Liao, QH Ji, QY Lei, W Hu
    Cell Reports, 2022;39(9):110851.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. A Factor H-Fc fusion protein increases complement-mediated opsonophagocytosis and killing of community associated methicillin-resistant Staphylococcus aureus
    Authors: MAG Sage, KD Cranmer, ML Semeraro, S Ma, EV Galkina, Y Tran, KL Wycoff, JA Sharp
    PLoS ONE, 2022;17(3):e0265774.
    Species: Human
    Sample Types: Serum
  5. Aggregated neutrophil extracellular traps occlude Meibomian glands during ocular surface inflammation
    Authors: A Mahajan, L Hasíková, U Hampel, A Grüneboom, X Shan, I Herrmann, F Garreis, F Bock, J Knopf, J Singh, C Schauer, S Mahajan, M Leppkes, F Paulsen, U Schlötzer-, V Krenn, A Jünemann, B Hohberger, G Schett, M Herrmann, LE Muñoz
    The ocular surface, 2021;20(0):1-12.
    Species: Human
    Sample Types: Tears
  6. Tissue-specific endothelial cell heterogeneity contributes to unequal inflammatory responses
    Authors: H Gunawardan, T Romero, N Yao, S Heidt, A Mulder, DA Elashoff, NM Valenzuela
    Scientific Reports, 2021;11(1):1949.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Therapeutic Hypothermia Inhibits the Classical Complement Pathway in a Rat Model of Neonatal Hypoxic-Ischemic Encephalopathy
    Authors: TA Shah, HK Pallera, CL Kaszowski, WT Bass, FA Lattanzio
    Frontiers in Neuroscience, 2021;15(0):616734.
    Species: Rat
    Sample Types: Plasma
  8. Radiotherapy-Induced Changes in the Systemic Immune and Inflammation Parameters of Head and Neck Cancer Patients
    Authors: K Balázs, E Kis, C Badie, EN Bogdándi, S Candéias, LC Garcia, I Dominczyk, B Frey, U Gaipl, Z Jurányi, ZS Kocsis, EA Rutten, G Sáfrány, P Widlak, K Lumniczky
    Cancers (Basel), 2019;11(9):.
    Species: Human
    Sample Types: Plasma
  9. Frontline Science: Aggregated neutrophil extracellular traps prevent inflammation on the neutrophil-rich ocular surface
    Authors: A Mahajan, A Grüneboom, L Petru, MJ Podolska, L Kling, C Maueröder, F Dahms, S Christians, L Günter, V Krenn, A Jünemann, F Bock, C Schauer, G Schett, B Hohberger, M Herrmann, LE Muñoz
    J. Leukoc. Biol., 2019;0(0):.
    Species: Human
    Sample Types: Eye Tear Fluid
  10. Broad adsorption of sepsis-related PAMP and DAMP molecules, mycotoxins, and cytokines from whole blood using CytoSorb� sorbent porous polymer beads
    Authors: MC Gruda, KG Ruggeberg, P O'Sullivan, T Guliashvil, AR Scheirer, TD Golobish, VJ Capponi, PP Chan
    PLoS ONE, 2018;13(1):e0191676.
    Species: Human
    Sample Types: Complex Sample Type
  11. 3D artificial round section micro-vessels to investigate endothelial cells under physiological flow conditions
    Authors: R Sfriso, S Zhang, CA Bichsel, O Steck, A Despont, OT Guenat, R Rieben
    Sci Rep, 2018;8(1):5898.
    Species: Human
    Sample Types: Serum
  12. Surface modification of pig endothelial cells with a branched heparin conjugate improves their compatibility with human blood
    Authors: AK Bongoni, E Salvaris, S Nordling, N Klymiuk, E Wolf, DL Ayares, R Rieben, PU Magnusson, PJ Cowan
    Sci Rep, 2017;7(1):4450.
    Species: Human
    Sample Types: Plasma
  13. TLR7/8 activation in neutrophils impairs immune complex phagocytosis through shedding of FcgRIIA
    Authors: C Lood, S Arve, J Ledbetter, KB Elkon
    J. Exp. Med., 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Complement effectors, C5a and C3a, in cystic fibrosis lung fluid correlate with disease severity
    Authors: PS Hair, LA Sass, T Vazifedan, TA Shah, NK Krishna, KM Cunnion
    PLoS ONE, 2017;12(3):e0173257.
    Species: Human
    Sample Types: Sputum
  15. Peripheral trafficking of bone-marrow-derived stem cells in patients with different types of gastric neoplasms
    Authors: W B?ogowski, E Zuba-Surma, D Sa?ata, M Budkowska, B Do??gowska, T Starzy?ska
    Oncoimmunology, 2016;5(4):e1099798.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
    Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
    PLoS ONE, 2015;10(8):e0134619.
    Species: Human
    Sample Types: Plasma
  17. Zinc supplementation inhibits complement activation in age-related macular degeneration.
    Authors: Smailhodzic D, van Asten F, Blom A, Mohlin F, den Hollander A, van de Ven J, van Huet R, Groenewoud J, Tian Y, Berendschot T, Lechanteur Y, Fauser S, de Bruijn C, Daha M, van der Wilt G, Hoyng C, Klevering B
    PLoS ONE, 2014;9(11):e112682.
    Species: Human
    Sample Types: Serum
  18. Clinical hypothermia temperatures increase complement activation and cell destruction via the classical pathway.
    Authors: Shah T, Mauriello C, Hair P, Sandhu A, Stolz M, Bass W, Krishna N, Cunnion K
    J Transl Med, 2014;12(0):181.
    Species: Human
    Sample Types: Serum
  19. Pulp fibroblasts synthesize functional complement proteins involved in initiating dentin-pulp regeneration.
    Authors: Chmilewsky F, Jeanneau C, Laurent P, About I
    Am J Pathol, 2014;184(7):1991-2000.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Citrate confers less filter-induced complement activation and neutrophil degranulation than heparin when used for anticoagulation during continuous venovenous haemofiltration in critically ill patients.
    Authors: Schilder L, Nurmohamed S, ter Wee P, Paauw N, Girbes A, Beishuizen A, Beelen R, Groeneveld A
    BMC Nephrol, 2014;15(0):19.
    Species: Human
    Sample Types: Serum
  21. Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
    Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
    BMC Infect Dis, 2014;14(0):35.
    Species: Human
    Sample Types: Serum
  22. Staphylococcus aureus surface protein SdrE binds complement regulator factor H as an immune evasion tactic.
    Authors: Sharp JA, Echague CG, Hair PS
    PLoS ONE, 2012;7(5):e38407.
    Species: Human
    Sample Types: Cell Culture Supernates


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Human Complement Component C5a DuoSet ELISA
By Anonymous on 09/14/2019
Sample Tested: Serum and Plasma

Human Complement Component C5a DuoSet ELISA
By Anonymous on 11/29/2017
Sample Tested: Serum