Mouse Complement Component C5a DuoSet ELISA

Catalog # Availability Size / Price Qty
DY2150
Ancillary Products Available
Mouse Complement Component C5a ELISA Standard Curve
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Product Details
Procedure
Citations (29)
FAQs
Supplemental Products
Reviews (2)

Mouse Complement Component C5a DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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 mouse 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)

 

  

Data Example

Mouse Complement Component C5a ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
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

Mouse 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 77 amino acid (aa) peptide that is created by the activity of C5a convertase on the C5 alpha -chain. Mouse C5a has four alpha -helices, plus three intrachain disulfide bonds that create a triple loop structure. In serum, proteolytic processing removes the C-terminal arginine, creating a low activity C5a desArg77 molecule. Mouse C5a shares 60% and 82% aa sequence identity to human and rat C5a, respectively. C5a binds to a signaling G-protein coupled receptor (GPCR) (C5aR/CD88), inducing neutrophil chemotaxis and endothelial cell activation. It also triggers an oxidative burst in macrophages and neutrophils, and induces release of histamine in basophils and mast cells. Alternatively, it may also bind to a nonsignaling GPCR termed C5L2 whose function is yet to be determined.

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

Assay Procedure

GENERAL ELISA PROTOCOL

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 Mouse 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.

29 Citations: Showing 1 - 10
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  1. Complement receptor C3aR1 controls neutrophil mobilization following spinal cord injury through physiological antagonism of CXCR2
    Authors: FH Brennan, T Jogia, ER Gillespie, LV Blomster, XX Li, B Nowlan, GM Williams, E Jacobson, GW Osborne, FA Meunier, SM Taylor, KE Campbell, KP MacDonald, JP Levesque, TM Woodruff, MJ Ruitenberg
    JCI Insight, 2019;4(9):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. Plasma protein adsorption on Fe3O4-PEG nanoparticles activates the complement system and induces an inflammatory response
    Authors: V Escamilla-, A Solorio-Ro, M Uribe-Ramí, O Lozano, S Lucas, A Chagolla-L, R Winkler, A De Vizcaya
    Int J Nanomedicine, 2019;14(0):2055-2067.
    Species: Mouse
    Sample Types: Plasma
  3. rIgG1 Fc Hexamer Inhibits Antibody-Mediated Autoimmune Disease via Effects on Complement and Fc?Rs
    Authors: R Spirig, IK Campbell, S Koernig, CG Chen, BJB Lewis, R Butcher, I Muir, S Taylor, J Chia, D Leong, J Simmonds, P Scotney, P Schmidt, L Fabri, A Hofmann, M Jordi, MO Spycher, S Cattepoel, J Brasseit, C Panousis, T Rowe, DR Branch, A Baz Morell, F Käsermann, AW Zuercher
    J. Immunol., 2018;0(0):.
    Species: Mouse
    Sample Types: Serum
  4. Complement Inhibitor CRIg/FH Ameliorates Renal Ischemia Reperfusion Injury via Activation of PI3K/AKT Signaling
    Authors: C Hu, L Li, P Ding, L Li, X Ge, L Zheng, X Wang, J Wang, W Zhang, N Wang, H Gu, F Zhong, M Xu, R Rong, T Zhu, W Hu
    J. Immunol., 2018;0(0):.
    Species: Mouse
    Sample Types: Serum
  5. Harmful Roles of TLR3 and TLR9 in Cardiac Dysfunction Developing during Polymicrobial Sepsis
    Authors: F Fattahi, MW Russell, EA Malan, M Parlett, E Abe, FS Zetoune, PA Ward
    Biomed Res Int, 2018;2018(0):4302726.
    Species: Mouse
    Sample Types: Plasma
  6. Complement depletion with cobra venom factor alleviates acute hepatic injury induced by ischemia?reperfusion
    Authors: B Wang, H Xu, J Li, HM Gao, YH Xing, Z Lin, HJ Li, YQ Wang, SH Cao
    Mol Med Rep, 2018;0(0):.
    Species: Rat
    Sample Types: Serum
  7. Complement C5aR1 Signaling Promotes Polarization and Proliferation of Embryonic Neural Progenitor Cells through PKC?
    Authors: LG Coulthard, OA Hawksworth, R Li, A Balachandr, JD Lee, F Sepehrband, N Kurniawan, A Jeanes, DG Simmons, E Wolvetang, TM Woodruff
    J. Neurosci., 2017;0(0):.
    Species: Mouse
    Sample Types: CSF
  8. Osteoblast-specific overexpression of complement receptor C5aR1 impairs fracture healing
    Authors: S Bergdolt, A Kovtun, Y Hägele, A Liedert, T Schinke, M Amling, M Huber-Lang, A Ignatius
    PLoS ONE, 2017;12(6):e0179512.
    Species: Mouse
    Sample Types: Serum
  9. Complement Activation and STAT4 Expression Are Associated with Early Inflammation in Diabetic Wounds
    Authors: KM Cunnion, NK Krishna, HK Pallera, A Pineros-Fe, MG Rivera, PS Hair, BP Lassiter, R Huyck, MA Clements, AF Hood, GT Rodeheaver, PS Cottler, JL Nadler, AD Dobrian
    PLoS ONE, 2017;12(1):e0170500.
    Species: Mouse
    Sample Types: Tissue Secretion
  10. Chronic lung inflammation primes humoral immunity and augments antipneumococcal resistance
    Authors: JD Boehme, S Stegemann-, A Autengrube, N Peters, J Wissing, L Jänsch, A Jeron, D Bruder
    Sci Rep, 2017;7(1):4972.
    Species: Mouse
    Sample Types: BALF
  11. Overexpression of Human CD55 and CD59 or Treatment with Human CD55 Protects against Renal Ischemia-Reperfusion Injury in Mice
    Authors: AK Bongoni, B Lu, EJ Salvaris, V Roberts, D Fang, JL McRae, N Fisicaro, KM Dwyer, PJ Cowan
    J. Immunol., 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. TLR-Induced Murine Dendritic Cell (DC) Activation Requires DC-Intrinsic Complement
    Authors: JH Sheen, MG Strainic, J Liu, W Zhang, Z Yi, ME Medof, PS Heeger
    J. Immunol., 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. Attenuation of cGVHD by C5a/C5aR blockade is associated with increased frequency of Treg
    Authors: Y Wang, P Lai, X Chen, C He, X Huang, S Geng, C Luo, S Wu, W Ling, L Zhong, Z Lu, P Li, J Weng, X Du
    Sci Rep, 2017;7(1):3603.
    Species: Mouse
    Sample Types: Plasma
  14. High-fat diet-induced complement activation mediates intestinal inflammation and neoplasia, independent of obesity
    Mol Cancer Res, 2016;0(0):.
    Species: Mouse
    Sample Types: Plasma
  15. The Complement System Component C5a Produces Thermal Hyperalgesia via Macrophage-to-Nociceptor Signaling That Requires NGF and TRPV1
    J Neurosci, 2016;36(18):5055-70.
    Species: Mouse
    Sample Types: Tissue Homogenates
  16. Lysophosphatidic acid alters the expression profiles of angiogenic factors, cytokines, and chemokines in mouse liver sinusoidal endothelial cells.
    Authors: Chou C, Lai S, Ho C, Lin W, Chen C, Lee P, Peng F, Kuo S, Wu S, Lai H
    PLoS ONE, 2015;10(3):e0122060.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  17. Complement component C5 recruits neutrophils in the absence of C3 during respiratory infection with modified vaccinia virus Ankara.
    Authors: Price P, Banki Z, Scheideler A, Stoiber H, Verschoor A, Sutter G, Lehmann M
    J Immunol, 2015;194(3):1164-8.
    Species: Mouse
    Sample Types: BALF
  18. Real-Time Imaging of Interactions of Neutrophils with Cryptococcus neoformans Demonstrates a Crucial Role of Complement C5a-C5aR Signaling.
    Authors: Sun D, Zhang M, Liu G, Wu H, Zhu X, Zhou H, Shi M
    Infect Immun, 2015;84(1):216-29.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  19. The Complement Receptor C5aR Controls Acute Inflammation and Astrogliosis following Spinal Cord Injury.
    Authors: Brennan F, Gordon R, Lao H, Biggins P, Taylor S, Franklin R, Woodruff T, Ruitenberg M
    J Neurosci, 2015;35(16):6517-31.
    Species: Mouse
    Sample Types: Plasma
  20. The triggering receptor expressed on myeloid cells 2 inhibits complement component 1q effector mechanisms and exerts detrimental effects during pneumococcal pneumonia.
    Authors: Sharif O, Gawish R, Warszawska J, Martins R, Lakovits K, Hladik A, Doninger B, Brunner J, Korosec A, Schwarzenbacher R, Berg T, Kralovics R, Colinge J, Mesteri I, Gilfillan S, Salmaggi A, Verschoor A, Colonna M, Knapp S
    PLoS Pathog, 2014;10(6):e1004167.
    Species: Mouse
    Sample Types: BALF
  21. Complement C3 and C5 deficiency affects fracture healing.
    Authors: Ehrnthaller, Christia, Huber-Lang, Markus, Nilsson, Per, Bindl, Ronny, Redeker, Simon, Recknagel, Stefan, Rapp, Anna, Mollnes, Tom, Amling, Michael, Gebhard, Florian, Ignatius, Anita
    PLoS ONE, 2013;8(11):e81341.
    Species: Mouse
    Sample Types: Serum
  22. Deficiency of C5L2 increases macrophage infiltration and alters adipose tissue function in mice.
    Authors: Gauvreau D, Gupta A, Fisette A, Tom F, Cianflone K
    PLoS ONE, 2013;8(4):e60795.
    Species: Mouse
    Sample Types: Plasma
  23. Antigen-presenting cell-derived complement modulates graft-versus-host disease.
    Authors: Kwan WH, Hashimoto D, Paz-Artal E
    J. Clin. Invest., 2012;122(6):2234-8.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  24. A protective role for complement C3 protein during pandemic 2009 H1N1 and H5N1 influenza A virus infection.
    Authors: O'Brien KB, Morrison TE, Dundore DY
    PLoS ONE, 2011;6(3):e17377.
    Species: Mouse
    Sample Types: BALF
  25. Complement regulates CD4 T-cell help to CD8 T cells required for murine allograft rejection.
    Authors: Vieyra M, Leisman S, Raedler H, Kwan WH, Yang M, Strainic MG, Medof ME, Heeger PS
    Am. J. Pathol., 2011;179(2):766-74.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  26. Distinct Different Contributions of the Alternative and Classical Complement Activation Pathway for the Innate Host Response during Sepsis.
    Authors: Dahlke K, Wrann CD, Sommerfeld O, Sossdorf M, Recknagel P, Sachse S, Winter SW, Klos A, Stahl GL, Ma YX, Claus RA, Reinhart K, Bauer M, Riedemann NC
    J. Immunol., 2011;186(5):3066-3075.
    Species: Mouse
    Sample Types: Plasma
  27. The C5a Receptor on Mast Cells Is Critical for the Autoimmune Skin-blistering Disease Bullous Pemphigoid.
    Authors: Heimbach L, Li Z, Berkowitz P
    J. Biol. Chem., 2011;286(17):15003-9.
    Species: Mouse
    Sample Types: Tissue Homogenates
  28. Increased local concentration of complement C5a contributes to incisional pain in mice.
    Authors: Jang JH, Liang D, Kido K
    J Neuroinflammation, 2011;8(0):80.
    Species: Mouse
    Sample Types: Tissue Homogenates
  29. The absence of serum IgM enhances the susceptibility of mice to pulmonary challenge with Cryptococcus neoformans.
    Authors: Subramaniam KS, Datta K, Quintero E, Manix C, Marks MS, Pirofski LA
    J. Immunol., 2010;184(10):5755-67.
    Species: Mouse
    Sample Types: Tissue Homogenates

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Mouse Complement Component C5a DuoSet ELISA
By Anonymous on 08/10/2019
Application: Sample Tested: Cell Lysates

Mouse Complement Component C5a DuoSet ELISA
By Anonymous on 10/08/2018
Application: Sample Tested: Serum