Mouse CCL3/MIP-1 alpha DuoSet ELISA

Catalog # Availability Size / Price Qty
DY450
DY450-05
Ancillary Products Available
Mouse CCL3 / MIP-1 alpha ELISA Standard Curve
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Product Details
Procedure
Citations (33)
FAQs
Supplemental Products
Reviews (4)

Mouse CCL3/MIP-1 alpha DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For five or 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 CCL3/MIP-1alpha. 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 CCL3 / MIP-1 alpha 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: CCL3/MIP-1 alpha

MIP-1 alpha (macrophage inflammatory protein 1 alpha) is a member of the CC or beta chemokine subfamily that was originally purified from the conditioned media of an LPS-stimulated murine macrophage cell line. MIP-1 alpha acts as a chemoattractant to a variety of cells including monocytes, T cells, B cells and eosinophils.

The two human MIP-1 alpha genes arise by duplication/mutation. They code for MIP-1 alpha isoforms CCL3/LD78a and CCL3L1/LD78b, which share 94% amino acid sequence homology. Whereas the human CCL3/LD78a is a single-copy gene, the human CCL3L1/LD78b gene copy number varies within the population. Human CCL3L1/LD78b binds and signals through chemokine receptors CCR1, CCR5. When compared to CCL3/LD78a, CCL3L1/LD78b has higher binding affinity to CCR5, which also functions as a coreceptor for HIV-1 entry. The copy number of CCL3L1 is one of several genetic determinants of HIV-1 susceptibility.

Entrez Gene IDs:
6348 (Human); 20302 (Mouse); 25542 (Rat); 448787 (Canine)
Alternate Names:
C-C motif chemokine 3; MIP1-(a); AI323804; CCL3; chemokine (C-C motif) ligand 3; G0S19-1; LD78a; LD78alpha; MIP1 alpha; MIP-1 alpha; Mip1a; MIP-1alpha; MIP1-alpha; Scya3

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 CCL3/MIP-1 alpha 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.

33 Citations: Showing 1 - 10
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  1. New role of P2X7 receptor in an Alzheimer's disease mouse model
    Authors: E Martin, M Amar, C Dalle, I Youssef, C Boucher, C Le Duigou, M Brückner, A Prigent, V Sazdovitch, A Halle, JM Kanellopou, B Fontaine, B Delatour, C Delarasse
    Mol. Psychiatry, 2018;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Caspase-8 Acts in a Non-enzymatic Role as a Scaffold for Assembly of a Pro-inflammatory FADDosome Complex upon TRAIL Stimulation
    Authors: CM Henry, SJ Martin
    Mol. Cell, 2017;65(4):715-729.e5.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. Surface layer proteins from virulent Clostridium difficile ribotypes exhibit signatures of positive selection with consequences for innate immune response
    Authors: M Lynch, TA Walsh, I Marszalows, AE Webb, M MacAogain, TR Rogers, H Windle, D Kelleher, MJ O'Connell, CE Loscher
    BMC Evol. Biol, 2017;17(1):90.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Deletion of the Chemokine Binding Protein Gene from the Parapoxvirus Orf Virus Reduces Virulence and Pathogenesis in Sheep
    Authors: SB Fleming, C McCaughan, Z Lateef, A Dunn, LM Wise, NC Real, AA Mercer
    Front Microbiol, 2017;8(0):46.
    Species: Mouse
    Sample Types: Protein
  5. The Carbohydrate Lectin Receptor, Dectin-1, Mediates Immune Response to Exserohilum rostratum
    Infect. Immun, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. The transcription factor ATF7 mediates lipopolysaccharide-induced epigenetic changes in macrophages involved in innate immunological memory.
    Authors: Yoshida K, Maekawa T, Zhu Y, Renard-Guillet C, Chatton B, Inoue K, Uchiyama T, Ishibashi K, Yamada T, Ohno N, Shirahige K, Okada-Hatakeyama M, Ishii S
    Nat Immunol, 2015;16(10):1034-43.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. C1q Modulates the Response to TLR7 Stimulation by Pristane-Primed Macrophages: Implications for Pristane-Induced Lupus.
    Authors: Carlucci F, Ishaque A, Ling G, Szajna M, Sandison A, Donatien P, Cook H, Botto M
    J Immunol, 2015;196(4):1488-94.
    Species: Mouse
    Sample Types: Serum
  8. A critical role for the TLR signaling adapter Mal in alveolar macrophage-mediated protection against Bordetella pertussis.
    Authors: Bernard N, Finlay C, Tannahill G, Cassidy J, O'Neill L, Mills K
    Mucosal Immunol, 2015;8(5):982-92.
    Species: Mouse
    Sample Types: Tissue Homogenates
  9. Ethanolic Echinacea purpurea Extracts Contain a Mixture of Cytokine-Suppressive and Cytokine-Inducing Compounds, Including Some That Originate from Endophytic Bacteria.
    Authors: Todd D, Gulledge T, Britton E, Oberhofer M, Leyte-Lugo M, Moody A, Shymanovich T, Grubbs L, Juzumaite M, Graf T, Oberlies N, Faeth S, Laster S, Cech N
    PLoS ONE, 2015;10(5):e0124276.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Murine lung injury caused by Leptospira interrogans glycolipoprotein, a specific Na/K-ATPase inhibitor.
    Authors: Goncalves-de-Albuquerque C, Burth P, Silva A, de Moraes I, Oliveira F, Santelli R, Freire A, de Lima G, da Silva E, da Silva C, Morandi V, Bozza P, Younes-Ibrahim M, de Castro Faria Neto H, de Castro Faria M
    Respir Res, 2014;15(0):93.
    Species: Mouse
    Sample Types: BALF
  11. UVB-induced skin inflammation and cutaneous tissue injury is dependent on the MHC class I-like protein, CD1d.
    Authors: Ryser S, Schuppli M, Gauthier B, Hernandez D, Roye O, Hohl D, German B, Holzwarth J, Moodycliffe A
    J Invest Dermatol, 2014;134(1):192-202.
    Species: Mouse
    Sample Types: Tissue Homogenates
  12. Lack of platelet-activating factor receptor protects mice against diet-induced adipose inflammation and insulin-resistance despite fat pad expansion.
    Authors: Menezes-Garcia Z, Oliveira M, Lima R, Soriani F, Cisalpino D, Botion L, Teixeira M, Souza D, Ferreira A
    Obesity (Silver Spring), 2014;22(3):663-72.
    Species: Mouse
    Sample Types: Tissue Homogenates
  13. Intraperitoneal prophylaxis with CpG oligodeoxynucleotides protects neutropenic mice against intracerebral Escherichia coli K1 infection.
    Authors: Ribes S, Meister T, Ott M, Redlich S, Janova H, Hanisch U, Nessler S, Nau R
    J Neuroinflammation, 2014;11(0):14.
  14. A novel murine model of rhinoscleroma identifies Mikulicz cells, the disease signature, as IL-10 dependent derivatives of inflammatory monocytes.
    Authors: Fevre C, Almeida A, Taront S, Pedron T, Huerre M, Prevost M, Kieusseian A, Cumano A, Brisse S, Sansonetti P, Tournebize R
    EMBO Mol Med, 2013;5(4):516-30.
    Species: Mouse
    Sample Types: Tissue Homogenates
  15. Syntaxin binding protein 1 is not required for allergic inflammation via IgE-mediated mast cell activation.
    Authors: Wu Z, MacNeil A, Berman J, Lin T
    PLoS ONE, 2013;8(3):e58560.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. Role of Toll interleukin-1 receptor (IL-1R) 8, a negative regulator of IL-1R/Toll-like receptor signaling, in resistance to acute Pseudomonas aeruginosa lung infection.
    Authors: Veliz Rodriguez T, Moalli F, Polentarutti N, Paroni M, Bonavita E, Anselmo A, Nebuloni M, Mantero S, Jaillon S, Bragonzi A, Mantovani A, Riva F, Garlanda C
    Infect. Immun., 2012;80(1):100-9.
    Species: Mouse
    Sample Types: Serum
  17. Inflammatory chemokine release of astrocytes in vitro is reduced by all-trans retinoic acid.
    Authors: van Neerven S, Regen T, Wolf D, Nemes A, Johann S, Beyer C, Hanisch UK, Mey J
    J. Neurochem., 2010;114(5):1511-26.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  18. Mucosal delivery of human papillomavirus pseudovirus-encapsidated plasmids improves the potency of DNA vaccination.
    Authors: Graham BS, Kines RC, Corbett KS, Nicewonger J, Roberts JN, Schiller JT, Buck CB
    Mucosal Immunol, 2010;3(5):475-86.
    Species: Mouse
    Sample Types: BALF
  19. Chorionic gonadotropin alleviates thioglycollate-induced peritonitis by affecting macrophage function.
    Authors: Wan H, Coppens JM, van Helden-Meeuwsen CG, Leenen PJ, Van Rooijen N, Khan NA, Kiekens RC, Benner R, Versnel MA
    J. Leukoc. Biol., 2009;86(2):361-70.
    Species: Mouse
    Sample Types: Peritoneal Lavage Fluid
  20. VAMP-8 segregates mast cell-preformed mediator exocytosis from cytokine trafficking pathways.
    Authors: Tiwari N, Wang CC, Brochetta C, Ke G, Vita F, Qi Z, Rivera J, Soranzo MR, Zabucchi G, Hong W, Blank U
    Blood, 2008;111(7):3665-74.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  21. Central role of MyD88-dependent dendritic cell maturation and proinflammatory cytokine production to control Brucella abortus infection.
    Authors: Macedo GC, Magnani DM, Carvalho NB, Bruna-Romero O, Gazzinelli RT, Oliveira SC
    J. Immunol., 2008;180(2):1080-7.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  22. gammadeltaT cell-mediated regulation of chemokine producing macrophages during Listeria monocytogenes infection-induced inflammation.
    Authors: Tramonti D, Rhodes K, Martin N, Dalton JE, Andrew E, Carding SR
    J. Pathol., 2008;216(2):262-70.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  23. CCL3-CCR5 axis regulates intratumoral accumulation of leukocytes and fibroblasts and promotes angiogenesis in murine lung metastasis process.
    Authors: Wu Y, Li YY, Matsushima K, Baba T, Mukaida N
    J. Immunol., 2008;181(9):6384-93.
    Species: Mouse
    Sample Types: Tissue Homogenates
  24. Nod1-mediated innate immune recognition of peptidoglycan contributes to the onset of adaptive immunity.
    Authors: Fritz JH, Le Bourhis L, Sellge G, Magalhaes JG, Fsihi H, Kufer TA, Collins C, Viala J, Ferrero RL, Girardin SE, Philpott DJ
    Immunity, 2007;26(4):445-59.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  25. CD8+ T Cells are required for inflammation and destruction in cigarette smoke-induced emphysema in mice.
    Authors: Maeno T, Houghton AM, Quintero PA, Grumelli S, Owen CA, Shapiro SD
    J. Immunol., 2007;178(12):8090-6.
    Species: Mouse
    Sample Types: BALF
  26. Mycobacterium tuberculosis antigens specifically modulate CCR2 and MCP-1/CCL2 on lymphoid cells from human pulmonary hilar lymph nodes.
    Authors: Arias MA, Jaramillo G, Lopez YP, Mejia N, Mejia C, Pantoja AE, Shattock RJ, Garcia LF, Griffin GE
    J. Immunol., 2007;179(12):8381-91.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Effects of dexamethazone on LPS-induced activationand migration of mouse dendritic cells revealed by a genome-wide transcriptional analysis.
    Authors: Vizzardelli C, Pavelka N, Luchini A, Zanoni I, Bendickson L, Pelizzola M, Beretta O, Foti M, Granucci F, Nilsen-Hamilton M, Ricciardi-Castagnoli P
    Eur. J. Immunol., 2006;36(6):1504-15.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  28. Evidence for the opposing roles of different gamma delta T cell subsets in macrophage homeostasis.
    Authors: Tramonti D, Andrew EM, Rhodes K, Newton DJ, Carding SR
    Eur. J. Immunol., 2006;36(7):1729-38.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  29. Mucosal administration of CpG oligodeoxynucleotide elicits strong CC and CXC chemokine responses in the vagina and serves as a potent Th1-tilting adjuvant for recombinant gD2 protein vaccination against genital herpes.
    Authors: Tengvall S, Lundqvist A, Eisenberg RJ, Cohen GH, Harandi AM
    J. Virol., 2006;80(11):5283-91.
    Species: Mouse
    Sample Types: Tissue Homogenates
  30. Differential involvement of TLR2 and TLR4 in host survival during pulmonary infection with Chlamydia pneumoniae.
    Authors: Rodriguez N, Wantia N, Fend F, Durr S, Wagner H, Miethke T
    Eur. J. Immunol., 2006;36(5):1145-55.
    Species: Mouse
    Sample Types: Tissue Homogenates
  31. Immunosuppressive effects of CCL17 on pulmonary antifungal responses during pulmonary invasive aspergillosis.
    Authors: Carpenter KJ, Hogaboam CM
    Infect. Immun., 2005;73(11):7198-207.
    Species: Mouse
    Sample Types: Tissue Homogenates
  32. Specific engagement of TLR4 or TLR3 does not lead to IFN-beta-mediated innate signal amplification and STAT1 phosphorylation in resident murine alveolar macrophages.
    Authors: Punturieri A, Alviani RS, Polak T, Copper P, Sonstein J, Curtis JL
    J. Immunol., 2004;173(2):1033-42.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  33. MIP-1 gamma promotes receptor-activator-of-NF-kappa-B-ligand-induced osteoclast formation and survival.
    Authors: Okamatsu Y, Battaglino R, Spate U, Stashenko P
    J. Immunol., 2004;173(3):2084-90.
    Species: Mouse
    Sample Types: Whole Cells

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Mouse CCL3/MIP-1 alpha DuoSet ELISA
By Anonymous on 04/15/2019
Sample Tested: peritoneal lavage fluid

Mice (HIII and LIII strains) injected i.p. with 2,4,6,10 tetramethylpentadecane (pristane).


Mouse CCL3/MIP-1 alpha DuoSet ELISA
By Anonymous on 03/28/2018
Sample Tested: BMDC

Mouse CCL3/MIP-1 alpha DuoSet ELISA
By Anonymous on 10/06/2016
Sample Tested: Cell culture supernatant

Mouse CCL3/MIP-1 alpha DuoSet ELISA, 15 Plate
By Anonymous on 04/05/2016
Sample Tested: murine lung homogenate supernatant

homogenate was filtered through 0.22um filter before use, diluted 1:3 for analysis