Human CCL4/MIP-1 beta Quantikine ELISA Kit

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Human CCL4/MIP-1 beta ELISA Cell Culture Supernate Standard Curve
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Human CCL4/MIP-1 beta Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.0 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (10 uL), Serum (150 uL), EDTA Plasma (150 uL), Heparin Plasma (150 uL), Citrate Plasma (150 uL)
Sensitivity
11 pg/mL
Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates), 31.2 - 2,000 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Specificity
Natural and recombinant human MIP-1 beta
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.

Product Summary

The Quantikine Human MIP-1 beta Immunoassay is a 3.0 or 4.5 hour solid phase ELISA designed to measure MIP-1 beta in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-1 beta (Act-2 variant) and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant human MIP-1 beta. Results obtained using natural human MIP-1 beta 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 for natural human MIP-1 beta.

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
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 23.3 98.8 428 31.4 118 476
Standard Deviation 1.1 3.8 17.3 3.7 9.9 28
CV% 4.7 3.8 4 11.8 8.4 5.9

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 51.3 208 932 58.8 220 949
Standard Deviation 4.6 7.5 30.2 5.7 15.6 70.5
CV% 9 3.6 3.2 9.7 7.1 7.4

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 107 81-117
Citrate Plasma (n=5) 99 87-116
EDTA Plasma (n=5) 100 84-112
Heparin Plasma (n=5) 99 90-112
Serum (n=5) 99 88-113

Linearity

To assess the linearity of the assay, five samples were spiked with high concentrations of MIP-1 beta in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CCL4/MIP-1 beta ELISA Linearity

Data Examples

Human CCL4/MIP-1 beta ELISA Cell Culture Supernate Standard Curve

Human CCL4/MIP-1 beta ELISA Serum/Plasma Standard Curve

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CCL4/MIP-1 beta

CCL4/MIP-1 beta is a beta chemokine that is secreted at sites of inflammation by activated leukocytes, lymphocytes, vascular endothelial cells, and pulmonary smooth muscle cells. It attracts a variety of immune cells to sites of microbial infection as well as to other pathologic inflammation such as allergic asthma and ischemic myocardium. CCL4 is secreted from activated monocytes as a heterodimer with CCL3/MIP-1 alpha. It signals through CCR5, and an N-terminally trimmed form additionally interacts with CCR1 and CCR2. In humans, the ability of CCL4 to bind CCR5 inhibits the cellular entry of M-tropic HIV-1 which utilizes CCR5 as a coreceptor.

Entrez Gene IDs:
6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine)
Alternate Names:
ACT2; ACT-2; AT744.1; C-C motif chemokine 4; CCL4; chemokine (C-C motif) ligand 4; Exodus-3; G-26 T-lymphocyte-secreted protein; G-26; HC21; LAG1; LAG-1; Lymphocyte activation gene 1 protein; Macrophage inflammatory protein 1-beta; MIP1 beta; MIP-1 beta; MIP1B; MIP1B1; MIP-1-beta; MIP-1-beta(1-69); PAT 744; Protein H400; SCYA2; SCYA4; secreted protein G-26; SIS-gamma; small inducible cytokine A4 (homologous to mouse Mip-1b); Small-inducible cytokine A4; T-cell activation protein 2
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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. Add Assay Diluent
  4.   For Serum & Plasma Samples: Add 100 µL of Assay Diluent to each well.
    For Cell Culture Supernate Samples: Add 50 µL Assay Diluent to each well.

  5. Add Standard, Control, or Sample
  6.   For Serum & Plasma Samples: Add 150 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    For Cell Culture Supernate Samples: Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1.25 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 1.25 hours.
  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 for Human CCL4/MIP-1 beta Quantikine ELISA Kit

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.

39 Citations: Showing 1 - 10
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  1. Profiling of cytokines, chemokines and other soluble proteins as a potential biomarker in colorectal cancer and polyps
    Authors: NA Johdi, L Mazlan, I Sagap, R Jamal
    Cytokine, 2017;99(0):35-42.
    Species: Human
    Sample Types: Serum
  2. A novel orally available Syk/Src/Jak2 inhibitor, SKLB-850, showed potent anti-tumor activities in B cell lymphoma (BCL) models
    Authors: N Zhang, G Zhang, N Liu, W Lin, S Ji, M Zheng, K Chen, X Liang, G Li, Y Ma, J Zhu, T Niu, LL Li, J Li, YQ Wei, SY Yang
    Oncotarget, 2017;8(67):111495-111507.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Stability of cytokines, chemokines and soluble activation markers in unprocessed blood stored under different conditions
    Cytokine, 2016;84(0):17-24.
    Species: Human
    Sample Types: Serum
  4. Functional Differences between IgM and IgD Signaling in Chronic Lymphocytic Leukemia
    J Immunol, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Serum levels of chemokines CCL4 and CCL5 in cirrhotic patients indicate the presence of hepatocellular carcinoma.
    Authors: Sadeghi M, Lahdou I, Oweira H, Daniel V, Terness P, Schmidt J, Weiss K, Longerich T, Schemmer P, Opelz G, Mehrabi A
    Br J Cancer, 2015;113(5):756-62.
    Species: Human
    Sample Types: Serum
  6. 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 Types: Cell Culture Supernates
  7. Pleurocidin, a novel antimicrobial peptide, induces human mast cell activation through the FPRL1 receptor.
    Authors: Pundir P, Catalli A, Leggiadro C, Douglas S, Kulka M
    Mucosal Immunol, 2014;7(1):177-87.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Regulation of inflammatory chemokine receptors on blood T cells associated to the circulating versus liver chemokines in dengue Fever.
    Authors: de-Oliveira-Pinto LM, Marinho CF, Povoa TF, de Azeredo EL, de Souza LA, Barbosa LD, Motta-Castro AR, Alves AM, Avila CA, de Souza LJ, da Cunha RV, Damasco PV, Paes MV, Kubelka CF
    PLoS ONE, 2012;7(7):e38527.
    Species: Human
    Sample Types: Plasma
  9. 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 Types: Cell Culture Supernates
  10. 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 Types: Plasma
  11. Lenalidomide as initial therapy of elderly patients with chronic lymphocytic leukemia.
    Authors: Badoux XC, Keating MJ, Wen S
    Blood, 2011;118(13):3489-98.
    Species: Human
    Sample Types: Plasma
  12. 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 Types: Plasma
  13. Macrophage inflammatory protein-1beta induced cell adhesion with increased intracellular reactive oxygen species.
    Authors: Tatara Y, Ohishi M, Yamamoto K, Shiota A, Hayashi N, Iwamoto Y, Takeda M, Takagi T, Katsuya T, Ogihara T, Rakugi H
    J. Mol. Cell. Cardiol., 2009;47(1):104-11.
    Species: Human
    Sample Types: Plasma
  14. 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 Types: Cell Culture Supernates
  15. NF-kappaB modulates sensitivity to apoptosis, proinflammatory and migratory potential in short- versus long-term cultured human gammadelta lymphocytes.
    Authors: Ferrarini M, Delfanti F, Gianolini M, Rizzi C, Alfano M, Lazzarin A, Biswas P
    J. Immunol., 2008;181(9):5857-64.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Infiltrated neutrophils acquire novel chemokine receptor expression and chemokine responsiveness in chronic inflammatory lung diseases.
    Authors: Hartl D, Krauss-Etschmann S, Koller B, Hordijk PL, Kuijpers TW, Hoffmann F, Hector A, Eber E, Marcos V, Bittmann I, Eickelberg O, Griese M, Roos D
    J. Immunol., 2008;181(11):8053-67.
    Species: Human
    Sample Types: BALF
  17. 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 Types: Cell Culture Supernates
  18. 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 Types: Plasma
  19. High T-cell response to human cytomegalovirus induces chemokine-mediated endothelial cell damage.
    Authors: Bolovan-Fritts CA, Trout RN, Spector SA
    Blood, 2007;110(6):1857-63.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Endothelial damage from cytomegalovirus-specific host immune response can be prevented by targeted disruption of fractalkine-CX3CR1 interaction.
    Authors: Bolovan-Fritts CA, Spector SA
    Blood, 2007;111(1):175-82.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. A network-based analysis of the late-phase reaction of the skin.
    Authors: Benson M, Langston MA, Adner M, Andersson B, Torinssson-Naluai A, Cardell LO
    J. Allergy Clin. Immunol., 2006;118(1):220-5.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. 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 Types: Cell Culture Supernates
  23. Respiratory syncytial virus and other respiratory viruses during the first 3 months of life promote a local TH2-like response.
    Authors: Kristjansson S, Bjarnarson SP, Wennergren G, Palsdottir AH, Arnadottir T, Haraldsson A, Jonsdottir I
    J. Allergy Clin. Immunol., 2005;116(4):805-11.
    Species: Human
    Sample Types: Tissue Secretion
  24. 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 Types: BALF
  25. 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 Types: Cell Culture Supernates
  26. 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 Types: CSF
  27. Virally stimulated plasmacytoid dendritic cells produce chemokines and induce migration of T and NK cells.
    Authors: Megjugorac NJ, Young HA, Amrute SB, Olshalsky SL, Fitzgerald-Bocarsly P
    J. Leukoc. Biol., 2004;75(3):504-14.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. 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 Types: Cell Culture Supernates
  29. 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 Types: Plasma
  30. 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 Types: Cell Culture Supernates
  31. 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 Types: Cell Culture Supernates
  32. 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 Types: Cell Culture Supernates
  33. Impact of NNRTI compared to PI-based highly active antiretroviral therapy on CCR5 receptor expression, beta-chemokines and IL-16 secretion in HIV-1 infection.
    Authors: Burton CT, Hardy GA, Sullivan AK, Nelson MR, Gazzard B, Gotch FM, Imami N
    Clin. Exp. Immunol., 2002;130(2):286-92.
    Species: Human
    Sample Types: Plasma
  34. Skewing of cytotoxic activity and chemokine production, but not of chemokine receptor expression, in human type-1/-2 gamma delta T lymphocytes.
    Authors: D&amp;amp;apos;Ambrosio D
    Eur. J. Immunol., 2002;32(10):2934-43.
    Species: Human
    Sample Types: Cell Culture Supernates
  35. 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 Types: Cell Culture Supernates
  36. 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 Types: Cell Culture Supernates
  37. 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 Types: Plasma
  38. 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 Types: Cell Culture Supernates
  39. 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 Types: Cell Culture Supernates

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Human CCL4/MIP-1 beta Quantikine ELISA Kit
By Anonymous on 02/27/2018
Sample Tested: 2120Ep human embryonal carcinoma cell line