Human CCL3/MIP-1 alpha Quantikine ELISA Kit

(41 citations)
(1 Review)
  
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours or 4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (10 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL), Citrate Plasma (200 uL)
  • Sensitivity
    10 pg/mL
  • Assay Range
    31.2 - 1,000 pg/mL (Cell Culture Supernates), 46.9 - 1,500 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant human MIP-1 alpha
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC20, Quantikine Immunoassay Control Group 3 - Please Inquire
Product Summary
The Quantikine Human MIP-1 alpha Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure MIP-1 alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-1 alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring human MIP-1 alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of natural human MIP-1 alpha.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean11644272653.6313623
Standard Deviation3.35.9143.713.425.7
CV%2.81.31.96.94.34.1

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean140688108450.7306613
Standard Deviation12.560.455.45.915.137.8
CV%8.98.85.111.64.96.2

Recovery

The recovery of MIP-1 alpha spiked to different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 100 88-117
Citrate Plasma (n=5) 94 85-104
EDTA Plasma (n=5) 96 81-109
Heparin Plasma (n=5) 98 82-121
Serum (n=5) 95 87-113
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of MIP-1 alpha in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CCL3/MIP-1 alpha Quantikine ELISA Kit
Preparation and Storage
  • 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;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. For Serum & Plasma Samples Only: Add 50 µL Assay Diluent
    4.   For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.

    5. 200 µL Standard, Control, or Sample
    6.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well.
    10.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
      For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    11.   Aspirate and wash 3 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

    14. 50 µL Stop Solution
    15. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    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.

    41 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Functional Differences between IgM and IgD Signaling in Chronic Lymphocytic Leukemia
      J Immunol, 2016;0(0):.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. The phosphoinositide-3-kinase (PI3K)-delta and gamma inhibitor, IPI-145 (Duvelisib), overcomes signals from the PI3K/AKT/S6 pathway and promotes apoptosis in CLL.
      Authors: Balakrishnan K, Peluso M, Fu M, Rosin N, Burger J, Wierda W, Keating M, Faia K, O'Brien S, Kutok J, Gandhi V
      Leukemia, 2015;29(9):1811-22.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Low concentrations of recombinant granulocyte macrophage-colony stimulating factor derived from Chinese hamster ovary cells augments long-term bioactivity with delayed clearance in vitro.
      Authors: Hashimoto A, Tanaka T, Itoh Y, Yamagata A, Kitamura N, Tazawa R, Nakagaki K, Nakata K
      Cytokine, 2014;68(2):118-26.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. The Bruton tyrosine kinase inhibitor PCI-32765 thwarts chronic lymphocytic leukemia cell survival and tissue homing in vitro and in vivo.
      Authors: Ponader S, Chen SS, Buggy JJ
      Blood, 2012;119(5):1182-9.
      Species: Human
      Sample Type: Plasma
    5. Alveolar fibrocyte percentage is an independent predictor of poor outcome in patients with acute lung injury.
      Authors: Quesnel C, Piednoir P, Gelly J, Nardelli L, Garnier M, Lecon V, Lasocki S, Bouadma L, Philip I, Elbim C, Mentre F, Soler P, Crestani B, Dehoux M
      Crit. Care Med., 2012;40(1):21-8.
      Species: Human
      Sample Type: BALF
    6. Respiratory syncytial virus glycoprotein G interacts with DC-SIGN and L-SIGN to activate ERK1 and ERK2.
      Authors: Johnson TR, McLellan JS, Graham BS
      J. Virol., 2012;86(3):1339-47.
      Species: Human
      Sample Type: Cell Culture Supernates
    7. CCL3 (MIP-1alpha) plasma levels and the risk for disease progression in chronic lymphocytic leukemia.
      Authors: Sivina M, Hartmann E, Kipps TJ, Rassenti L, Krupnik D, Lerner S, LaPushin R, Xiao L, Huang X, Werner L, Neuberg D, Kantarjian H, O'Brien S, Wierda WG, Keating MJ, Rosenwald A, Burger JA
      Blood, 2011;117(5):1662-9.
      Species: Human
      Sample Type: Plasma
    8. A phase I trial of bortezomib with temozolomide in patients with advanced melanoma: toxicities, antitumor effects, and modulation of therapeutic targets.
      Authors: Su Y, Amiri KI, Horton LW
      Clin. Cancer Res., 2010;16(1):348-57.
      Species: Human
      Sample Type: Serum
    9. HIV increases markers of cardiovascular risk: results from a randomized, treatment interruption trial.
      Authors: Calmy A, Gayet-Ageron A, Montecucco F, Nguyen A, Mach F, Burger F, Ubolyam S, Carr A, Ruxungtham K, Hirschel B, Ananworanich J,
      AIDS, 2009;23(8):929-39.
      Species: Human
      Sample Type: Plasma
    10. Interleukin-32 promotes osteoclast differentiation but not osteoclast activation.
      Authors: Mabilleau G, Sabokbar A
      PLoS ONE, 2009;4(1):e4173.
      Species: Human
      Sample Type: Cell Culture Supernates
    11. Chronically inflamed human tissues are infiltrated by highly differentiated Th17 lymphocytes.
      Authors: Pene J, Chevalier S, Preisser L, Venereau E, Guilleux MH, Ghannam S, Moles JP, Danger Y, Ravon E, Lesaux S, Yssel H, Gascan H
      J. Immunol., 2008;180(0):7423.
      Species: Human
      Sample Type: Cell Culture Supernates
    12. Expression of CCR5 receptors on Reed-Sternberg cells and Hodgkin lymphoma cell lines: involvement of CCL5/Rantes in tumor cell growth and microenvironmental interactions.
      Authors: Aldinucci D, Lorenzon D, Cattaruzza L, Pinto A, Gloghini A, Carbone A, Colombatti A
      Int. J. Cancer, 2008;122(4):769-76.
      Species: Human
      Sample Type: Cell Culture Supernates
    13. MMP-12 induces IL-8/CXCL8 secretion through EGFR and ERK1/2 activation in epithelial cells.
      Authors: Le Quement C, Guenon I, Gillon JY, Lagente V, Boichot E
      Am. J. Physiol. Lung Cell Mol. Physiol., 2008;294(6):L1076-84.
      Species: Human
      Sample Type: Cell Culture Supernates
    14. Expression of CXC and CC type of chemokines and its receptors in tuberculous and non-tuberculous effusions.
      Authors: Pokkali S, Das SD, R L
      Cytokine, 2008;41(3):307-14.
      Species: Human
      Sample Type: Serum
    15. In vitro induction of a dendritic cell phenotype in primary human acute myelogenous leukemia (AML) blasts alters the chemokine release profile and increases the levels of T cell chemotactic CCL17 and CCL22.
      Authors: Olsnes AM, Ryningen A, Ersvaer E, Bruserud Ø
      J. Interferon Cytokine Res., 2008;28(5):297-310.
      Species: Human
      Sample Type: Cell Culture Supernates
    16. Duffy antigen modifies the chemokine response in human endotoxemia.
      Authors: Mayr FB, Spiel AO, Leitner JM, Firbas C, Kliegel T, Jilma-Stohlawetz P, Derendorf H, Jilma B
      Crit. Care Med., 2008;36(1):159-65.
      Species: Human
      Sample Type: Plasma
    17. Protection against inflammation- and autoantibody-caused fetal loss by the chemokine decoy receptor D6.
      Authors: Martinez de la Torre Y, Buracchi C, Borroni EM, Dupor J, Bonecchi R, Nebuloni M, Pasqualini F, Doni A, Lauri E, Agostinis C, Bulla R, Cook DN, Haribabu B, Meroni P, Rukavina D, Vago L, Tedesco F, Vecchi A, Lira SA, Locati M, Mantovani A
      Proc. Natl. Acad. Sci. U.S.A., 2007;104(7):2319-24.
      Species: Human
      Sample Type: Cell Culture Supernates
    18. IL-18 is induced and IL-18 receptor alpha plays a critical role in the pathogenesis of cigarette smoke-induced pulmonary emphysema and inflammation.
      Authors: Kang MJ, Homer RJ, Gallo A, Lee CG, Crothers KA, Cho SJ, Rochester C, Cain H, Chupp G, Yoon HJ, Elias JA
      J. Immunol., 2007;178(3):1948-59.
      Species: Human
      Sample Type: BALF
    19. Modulation of eotaxin-3 (CCL26) in alveolar type II epithelial cells.
      Authors: Abonyo BO, Lebby KD, Tonry JH, Ahmad M, Heiman AS
      Cytokine, 2006;36(5):237-44.
      Species: Human
      Sample Type: Cell Culture Supernates
    20. Human Vgamma2Vdelta2 T cells contain cytoplasmic RANTES.
      Authors: Tikhonov I, Deetz CO, Paca R, Berg S, Lukyanenko V, Lim JK, Pauza CD
      Int. Immunol., 2006;18(8):1243-51.
      Species: Human
      Sample Type: Cell Lysates
    21. CCR5 expression and CC chemokine levels in idiopathic pulmonary fibrosis.
      Authors: Capelli A, Di Stefano A, Gnemmi I, Donner CF
      Eur. Respir. J., 2005;25(4):701-7.
      Species: Human
      Sample Type: BALF
    22. Interleukin-25-induced chemokines and interleukin-6 release from eosinophils is mediated by p38 mitogen-activated protein kinase, c-Jun N-terminal kinase, and nuclear factor-kappaB.
      Authors: Wong CK, Cheung PF, Ip WK, Lam CW
      Am. J. Respir. Cell Mol. Biol., 2005;33(2):186-94.
      Species: Human
      Sample Type: Cell Culture Supernates
    23. Up-regulation of XCR1 expression in rheumatoid joints.
      Authors: Wang CR, Liu MF, Huang YH, Chen HC
      Rheumatology (Oxford), 2004;43(5):569-73.
      Species: Human
      Sample Type: Serum
    24. Concentrations of macrophage inflammatory proteins MIP-1alpha and MIP-1beta and interleukin 8 (il-8) in lyme borreliosis.
      Authors: Grygorczuk S, Pancewicz S, Zajkowska J, Kondrusik M, Rwierzbinska R, Hermanowska-Szpakowicz T
      Infection, 2004;32(6):350-5.
      Species: Human
      Sample Type: CSF
    25. Role for cysteinyl leukotrienes in allergen-induced change in circulating dendritic cell number in asthma.
      Authors: Parameswaran K, Liang H, Fanat A, Watson R, Snider DP, O&amp;amp;apos;Byrne PM
      J. Allergy Clin. Immunol., 2004;114(1):73-9.
      Species: Human
      Sample Type: Sputum
    26. Enhanced monocyte chemoattractant protein-3/CC chemokine ligand-7 in usual interstitial pneumonia.
      Authors: Choi ES, Jakubzick C, Carpenter KJ, Kunkel SL, Evanoff H, Martinez FJ, Flaherty KR, Toews GB, Colby TV, Kazerooni EA, Gross BH, Travis WD, Hogaboam CM
      Am. J. Respir. Crit. Care Med., 2004;170(5):508-15.
      Species: Human
      Sample Type: Cell Culture Supernates
    27. Inhibition of human immunodeficiency virus type 1 replication by Z-100, an immunomodulator extracted from human-type tubercle bacilli, in macrophages.
      Authors: Emori Y, Ikeda T, Ohashi T, Masuda T, Kurimoto T, Takei M, Kannagi M
      J. Gen. Virol., 2004;85(0):2603-13.
      Species: Human
      Sample Type: Cell Culture Supernates
    28. Regulation of CCR5 expression and MIP-1alpha production in CD4+ T cells from patients with rheumatoid arthritis.
      Authors: Wang CR, Liu MF
      Clin. Exp. Immunol., 2003;132(2):371-8.
      Species: Human
      Sample Type: Cell Culture Supernates
    29. Simian immunodeficiency viruses with defective nef genes show increased susceptibility to the noncytotoxic antiviral activity of CD8+ lymphocytes.
      Authors: Binninger-Schinzel D, Norley S, Adler HS, Oberg HH, Kurth R
      Virology, 2002;294(1):209-21.
      Species: Human
      Sample Type: Cell Culture Supernates
    30. CCR5-binding chemokines modulate CXCL12 (SDF-1)-induced responses of progenitor B cells in human bone marrow through heterologous desensitization of the CXCR4 chemokine receptor.
      Authors: Honczarenko M, Le Y, Glodek AM, Silberstein LE
      Blood, 2002;100(7):2321-9.
      Species: Human
      Sample Type: Cell Culture Supernates
    31. Endogenously expressed nef uncouples cytokine and chemokine production from membrane phenotypic maturation in dendritic cells.
      Authors: Messmer D, Jacque JM, Santisteban C, Bristow C, Han SY, Villamide-Herrera L, Mehlhop E, Marx PA, Steinman RM, Gettie A, Pope M
      J. Immunol., 2002;169(8):4172-82.
      Species: Human
      Sample Type: Cell Culture Supernates
    32. Analysis of chemokines and chemokine receptor expression in ovarian cancer ascites.
      Authors: Milliken D, Scotton C, Raju S, Balkwill F, Wilson J
      Clin. Cancer Res., 2002;8(4):1108-14.
      Species: Human
      Sample Type: Plasma
    33. TNFalpha-induced macrophage chemokine secretion is more dependent on NF-kappaB expression than lipopolysaccharides-induced macrophage chemokine secretion.
      Authors: Ciesielski CJ, Andreakos E, Foxwell BM, Feldmann M
      Eur. J. Immunol., 2002;32(7):2037-45.
      Species: Human
      Sample Type: Cell Culture Supernates
    34. Distinct hematopoietic support by two human stromal cell lines.
      Authors: Loeuillet C, Bernard G, Remy-Martin J, Saas P, Herve P, Douay L, Chalmers D
      Exp. Hematol., 2001;29(6):736-45.
      Species: Human
      Sample Type: Cell Culture Supernates
    35. Il-6 and its soluble receptor orchestrate a temporal switch in the pattern of leukocyte recruitment seen during acute inflammation.
      Authors: Hurst SM, Wilkinson TS, McLoughlin RM, Jones S, Horiuchi S, Yamamoto N, Rose-John S, Fuller GM, Topley N, Jones SA
      Immunity, 2001;14(6):705-14.
      Species: Human
      Sample Type: Cell Culture Supernates
    36. Amyloid beta and amylin fibrils induce increases in proinflammatory cytokine and chemokine production by THP-1 cells and murine microglia.
      Authors: Yates SL, Burgess LH, Kocsis-Angle J, Antal JM, Dority MD, Embury PB, Piotrkowski AM, Brunden KR
      J. Neurochem., 2000;74(3):1017-25.
      Species: Human
      Sample Type: Cell Culture Supernates
    37. Chemoattractant factors in breast milk from allergic and nonallergic mothers.
      Authors: Bottcher MF, Jenmalm MC, Bjorksten B, Garofalo RP
      Pediatr. Res., 2000;47(5):592-7.
      Species: Human
      Sample Type: Serum
    38. Up-regulation of beta-chemokines and down-modulation of CCR5 co-receptors inhibit simian immunodeficiency virus transmission in non-human primates.
      Authors: Lehner T, Wang Y, Cranage M, Tao L, Mitchell E, Bravery C, Doyle C, Pratt K, Hall G, Dennis M, Villinger L, Bergmeier L
      Immunology, 2000;99(4):569-77.
      Species: Primate - Macaca mulatta (Rhesus Macaque)
      Sample Type: Cell Culture Supernates
    39. Immunization with live attenuated simian immunodeficiency virus induces strong type 1 T helper responses and beta-chemokine production.
      Authors: Gauduin MC, Glickman RL, Ahmad S, Yilma T, Johnson RP
      Proc. Natl. Acad. Sci. U.S.A., 1999;96(24):14031-6.
      Species: Primate
      Sample Type: Cell Culture Supernates
    40. Interleukin-12 induces sustained activation of multiple host inflammatory mediator systems in chimpanzees.
      Authors: Lauw FN, Dekkers PE, te Velde AA, Speelman P, Levi M, Kurimoto M, Hack CE, Van Deventer SJ, van der Poll T
      J. Infect. Dis., 1999;179(3):646-52.
      Species: Primate - Pan troglodytes (Chimpanzee)
      Sample Type: Plasma
    41. Protective mucosal immunity elicited by targeted iliac lymph node immunization with a subunit SIV envelope and core vaccine in macaques.
      Authors: Lehner T, Wang Y, Cranage M, Bergmeier LA, Mitchell E, Tao L, Hall G, Dennis M, Cook N, Brookes R, Klavinskis L, Jones I, Doyle C, Ward R
      Nat. Med., 1996;2(7):767-75.
      Species: Primate - Macaca mulatta (Rhesus Macaque)
      Sample Type: Cell Culture Supernates
    Expand to show all 41 Citations
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    Sample TestedCell culture supernatant

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