Mouse CCL4/MIP-1 beta Quantikine ELISA Kit

  (25 citations)     
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Assay Procedure
Citations (25)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (25 uL), EDTA Plasma (25 uL)
  • Sensitivity
    3 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant mouse 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 Mouse MIP-1 beta Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MIP-1 beta levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse MIP-1 beta and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse MIP-1 beta accurately. Results obtained using natural mouse MIP-1 beta showed dose response curves that were parallel to the standard curves obtained using the recombinant standards. These results indicate that this kit can be used to determine relative mass values for natural mouse 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, Serum, EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 23.1 73 234 23.7 77 245
Standard Deviation 2.2 4 5.4 1.7 3.4 12.5
CV% 9.5 5.5 2.3 7.2 4.4 5.1

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 104 86-118
EDTA Plasma (n=5) 99 87-106
Serum (n=5) 101 93-109
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse MIP-1 beta in each matrix were diluted with Calibrator Diluent and then assayed.
 CCL4/MIP-1 beta [HRP]
Product Datasheets

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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
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. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 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 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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.

25 Citations: Showing 1 - 10
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Species
Sample Type
  1. Behavioural traits propagate across generations via segregated iterative-somatic and gametic epigenetic mechanisms
    Nat Commun, 2016;7(0):11492.
    Species: Mouse
    Sample Type: Plasma
  2. CCL3 production by microglial cells modulates disease severity in murine models of retinal degeneration.
    Authors: Kohno H, Maeda T, Perusek L, Pearlman E, Maeda A
    J Immunol, 2014;192(8):3816-27.
    Species: Mouse
    Sample Type: Tissue Homogenates
  3. Cleavage of the T cell protein tyrosine phosphatase by the hepatitis C virus nonstructural 3/4A protease induces a Th1 to Th2 shift reversible by ribavirin therapy.
    Authors: Brenndorfer E, Brass A, Karthe J, Ahlen G, Bode J, Sallberg M
    J Immunol, 2014;192(4):1671-80.
    Species: Mouse
    Sample Type: Cell Lysates
  4. Potential roles of CCR5(+) CCR6(+) dendritic cells induced by nasal ovalbumin plus Flt3 ligand expressing adenovirus for mucosal IgA responses.
    Authors: Fukuyama Y, Tokuhara D, Sekine S, Aso K, Kataoka K, Davydova J, Yamamoto M, Gilbert R, Tokuhara Y, Fujihashi K, Kunisawa J, Yuki Y, Kiyono H, McGhee J, Fujihashi K
    PLoS ONE, 2013;8(4):e60453.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  5. Interferon gamma-induced intratumoral expression of CXCL9 alters the local distribution of T cells following immunotherapy with
    Authors: Guirnalda P, Wood L, Goenka R, Crespo J, Paterson Y
    Oncoimmunology, 2013;2(8):e25752.
    Species: Mouse
    Sample Type: Tissue Homogenates
  6. Modelling of mouse experimental colitis by global property screens: a holistic approach to assess drug effects in inflammatory bowel disease.
    Authors: Gottfries J, Melgar S, Michaelsson E
    PLoS ONE, 2012;7(1):e30005.
    Species: Mouse
    Sample Type: Tissue Homogenates
  7. Endotoxin augmented antigen-induced Th1 cell trafficking amplifies airway neutrophilic inflammation.
    Authors: Mikhak Z, Farsidjani A, Luster AD
    J. Immunol., 2009;182(12):7946-56.
    Species: Mouse
    Sample Type: BALF
  8. TLR9 agonist, but not TLR7/8, functions as an adjuvant to diminish FI-RSV vaccine-enhanced disease, while either agonist used as therapy during primary RSV infection increases disease severity.
    Authors: Johnson TR, Rao S, Seder RA, Chen M, Graham BS
    Vaccine, 2009;27(23):3045-52.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  9. CCR5 is involved in resolution of inflammation in proteoglycan-induced arthritis.
    Authors: Doodes PD, Cao Y, Hamel KM, Wang Y, Rodeghero RL, Kobezda T, Finnegan A
    Arthritis Rheum., 2009;60(10):2945-53.
    Species: Mouse
    Sample Type: Serum
  10. Pulmonary eosinophilia requires interleukin-5, eotaxin-1, and CD4+ T cells in mice immunized with respiratory syncytial virus G glycoprotein.
    Authors: Johnson TR, Rothenberg ME, Graham BS
    J. Leukoc. Biol., 2008;84(3):748-59.
    Species: Mouse
    Sample Type: Tissue Homogenates
  11. Role of the chemokine decoy receptor D6 in balancing inflammation, immune activation, and antimicrobial resistance in Mycobacterium tuberculosis infection.
    Authors: Di Liberto D, Locati M, Caccamo N, Vecchi A, Meraviglia S, Salerno A, Sireci G, Nebuloni M, Caceres N, Cardona PJ, Dieli F, Mantovani A
    J. Exp. Med., 2008;205(9):2075-84.
    Species: Mouse
    Sample Type: Serum
  12. Cannabinoid-2 receptor mediates protection against hepatic ischemia/reperfusion injury.
    Authors: Batkai S, Osei-Hyiaman D, Pan H, El-Assal O, Rajesh M, Mukhopadhyay P, Hong F, Harvey-White J, Jafri A, Hasko G, Huffman JW, Gao B, Kunos G, Pacher P
    FASEB J., 2007;21(8):1788-800.
    Species: Mouse
    Sample Type: Tissue Homogenates
  13. CCL4 protects from type 1 diabetes by altering islet beta-cell-targeted inflammatory responses.
    Authors: Meagher C, Arreaza G, Peters A, Strathdee CA, Gilbert PA, Mi QS, Santamaria P, Dekaban GA, Delovitch TL
    Diabetes, 2007;56(3):809-17.
    Species: Mouse
    Sample Type: Serum
  14. The direct action of 1alpha,25(OH)2-vitamin D3 on purified mouse Langerhans cells.
    Authors: Fujita H, Asahina A, Komine M, Tamaki K
    Cell. Immunol., 2007;245(2):70-9.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  15. Organ-specific role of MyD88 for gene regulation during polymicrobial peritonitis.
    Authors: Weighardt H, Mages J, Jusek G, Kaiser-Moore S, Lang R, Holzmann B
    Infect. Immun., 2006;74(6):3618-32.
    Species: Mouse
    Sample Type: Tissue Homogenates
  16. Immunosuppressant triptolide inhibits dendritic cell-mediated chemoattraction of neutrophils and T cells through inhibiting Stat3 phosphorylation and NF-kappaB activation.
    Authors: Liu Q, Chen T, Chen G, Li N, Wang J, Ma P, Cao X
    Biochem. Biophys. Res. Commun., 2006;345(3):1122-30.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  17. Deficiency of P-selectin or P-selectin glycoprotein ligand-1 leads to accelerated development of glomerulonephritis and increased expression of CC chemokine ligand 2 in lupus-prone mice.
    Authors: He X, Schoeb TR, Panoskaltsis-Mortari A, Zinn KR, Kesterson RA, Zhang J, Samuel S, Hicks MJ, Hickey MJ, Bullard DC
    J. Immunol., 2006;177(12):8748-56.
    Species: Mouse
    Sample Type: Tissue Homogenates
  18. Local production of chemokines and prostaglandin E2 in the acute, chronic and recovery phase of murine experimental colitis.
    Authors: Drmotova M, Rehnstrom E, Jansson L
    Cytokine, 2006;35(5):275-83.
    Species: Mouse
    Sample Type: Tissue Homogenates
  19. Immunization with gp120-depleted whole killed HIV immunogen and a second-generation CpG DNA elicits strong HIV-specific responses in mice.
    Authors: Trabattoni D, Clivio A, Bray DH, Bhagat L, Beltrami S, Maffeis G, Cesana E, Lowry P, Lissoni F, Kandimalla ER, Sullivan T, Agrawal S, Bartholomew R, Clerici M
    Vaccine, 2005;24(9):1470-7.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  20. The role of gammadelta T cells in the regulation of neutrophil-mediated tissue damage after thermal injury.
    Authors: Toth B, Alexander M, Daniel T, Hubbard WJ, Schwacha MG
    J. Leukoc. Biol., 2004;76(3):545-52.
    Species: Mouse
    Sample Type: Plasma
  21. Vbeta14(+) T cells mediate the vaccine-enhanced disease induced by immunization with respiratory syncytial virus (RSV) G glycoprotein but not with formalin-inactivated RSV.
    Authors: Johnson TR, Varga SM, Braciale TJ, Graham BS
    J. Virol., 2004;78(16):8753-60.
    Species: Mouse
    Sample Type: Tissue Homogenates
  22. Effects of poly(ADP-ribose) polymerase inhibition on inflammatory cell migration in a murine model of asthma.
    Authors: Virag L, Bai P, Bak I, Pacher P, Mabley JG, Liaudet L, Bakondi E, Gergely P, Kollai M, Szabo C
    Med. Sci. Monit., 2004;10(3):BR77-83.
    Species: Mouse
    Sample Type: BALF
  23. Role of the autocrine chemokines MIP-1alpha and MIP-1beta in the metastatic behavior of murine T cell lymphoma.
    Authors: Menten P, Saccani A, Dillen C, Wuyts A, Struyf S, Proost P, Mantovani A, Wang JM, Van Damme J
    J. Leukoc. Biol., 2002;72(4):780-9.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  24. Immunostimulatory DNA-based vaccines elicit multifaceted immune responses against HIV at systemic and mucosal sites.
    Authors: Horner AA, Datta SK, Takabayashi K, Belyakov IM, Cinman N, Nguyen MD, Van Uden JH, Berzofsky JA
    J. Immunol., 2001;167(3):1584-91.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  25. Inhibition of lipopolysaccharide-induced signal transduction in endotoxin-tolerized mouse macrophages: dysregulation of cytokine, chemokine, and toll-like receptor 2 and 4 gene expression.
    Authors: Medvedev AE, Kopydlowski KM, Vogel SN
    J. Immunol., 2000;164(11):5564-74.
    Species: Mouse
    Sample Type: Cell Culture Supernates
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