Human CCL3/MIP-1 alpha DuoSet ELISA

Catalog # Availability Size / Price Qty
DY270
DY270-05
Ancillary Products Available
Human CCL3 / MIP-1 alpha ELISA Standard Curve
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Product Details
Procedure
Citations (33)
FAQs
Supplemental Products
Reviews (3)

Human 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 human 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

Human 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); 102136134 (Cynomolgus Monkey)
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; PAT 464.1; SCYA3; SIS-beta

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 Human 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. Bone marrow-derived mesenchymal stem cells promote colorectal cancer progression via CCR5
    Authors: G Nishikawa, K Kawada, J Nakagawa, K Toda, R Ogawa, S Inamoto, R Mizuno, Y Itatani, Y Sakai
    Cell Death Dis, 2019;10(4):264.
  2. Serum Apolipoprotein E and Other Inflammatory Markers Can Identify Non-Responding Patients to a Dendritic Cell Vaccine
    Authors: H Leeman, E Kaminska, D Green, M Bodman-Smi, A Gravett, K Bodman-Smi, J Copier, G Coulton, A Fusi, AG Dalgleish
    Transl Oncol, 2019;12(3):397-403.
    Species: Human
    Sample Types: Serum
  3. Broad adsorption of sepsis-related PAMP and DAMP molecules, mycotoxins, and cytokines from whole blood using CytoSorb� sorbent porous polymer beads
    Authors: MC Gruda, KG Ruggeberg, P O'Sullivan, T Guliashvil, AR Scheirer, TD Golobish, VJ Capponi, PP Chan
    PLoS ONE, 2018;13(1):e0191676.
    Species: Human
    Sample Types: Complex Sample Type
  4. CUDC-907 blocks multiple pro-survival signals and abrogates microenvironment protection in CLL
    Authors: Y Chen, C Peubez, V Smith, S Xiong, G Kocsis-Fod, B Kennedy, S Wagner, C Balotis, S Jayne, MJS Dyer, S Macip
    J. Cell. Mol. Med., 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Altered distribution of peripheral blood dendritic cell subsets in patients with pulmonary paracoccidioidomycosis
    Authors: J Venturini, RS Cavalcante, DV Moris, M de Assis G, AD Levorato, KH Dos Reis, MSP de Arruda, RP Mendes
    Acta Trop., 2017;0(0):.
    Species: Human
    Sample Types: Serum
  6. Relevance of CCL3/CCR5 axis in oral carcinogenesis
    Authors: JM da Silva, TP Moreira Do, LM Sobral, CM Queiroz-Ju, MA Rachid, AEI Proudfoot, GP Garlet, AC Batista, MM Teixeira, AM Leopoldino, RC Russo, TA Silva
    Oncotarget, 2017;8(31):51024-51036.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Biocompatibility of primers and an adhesive used for implant-retained maxillofacial prostheses: An in�vitro analysis
    Authors: Daniela Micheline Dos Santos
    J Prosthet Dent, 2016;0(0):.
  8. CD84 mediates CLL-microenvironment interactions
    Oncogene, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. In vitro progesterone modulation on bacterial endotoxin-induced production of IL-1beta, TNFalpha, IL-6, IL-8, IL-10, MIP-1alpha, and MMP-9 in pre-labor human term placenta.
    Authors: Garcia-Ruiz G, Flores-Espinosa P, Preciado-Martinez E, Bermejo-Martinez L, Espejel-Nunez A, Estrada-Gutierrez G, Maida-Claros R, Flores-Pliego A, Zaga-Clavellina V
    Reprod Biol Endocrinol, 2015;13(0):115.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Neutrophil oxidative burst activates ATM to regulate cytokine production and apoptosis.
    Authors: Harbort C, Soeiro-Pereira P, von Bernuth H, Kaindl A, Costa-Carvalho B, Condino-Neto A, Reichenbach J, Roesler J, Zychlinsky A, Amulic B
    Blood, 2015;126(26):2842-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Human osteoarthritic cartilage shows reduced in vivo expression of IL-4, a chondroprotective cytokine that differentially modulates IL-1beta-stimulated production of chemokines and matrix-degrading enzymes in vitro.
    Authors: Assirelli E, Pulsatelli L, Dolzani P, Platano D, Olivotto E, Filardo G, Trisolino G, Facchini A, Borzi R, Meliconi R
    PLoS ONE, 2014;9(5):e96925.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Human B cells induce dendritic cell maturation and favour Th2 polarization by inducing OX-40 ligand.
    Authors: Maddur M, Sharma M, Hegde P, Stephen-Victor E, Pulendran B, Kaveri S, Bayry J
    Nat Commun, 2014;5(0):4092.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Thrombin-cleaved fragments of osteopontin are overexpressed in malignant glial tumors and provide a molecular niche with survival advantage.
    Authors: Yamaguchi Y, Shao Z, Sharif S, Du X, Myles T, Merchant M, Harsh G, Glantz M, Recht L, Morser J, Leung L
    J Biol Chem, 2013;288(5):3097-111.
    Species: Human
    Sample Types: CSF
  14. Selective activation of human dendritic cells by OM-85 through a NF-kB and MAPK dependent pathway.
    Authors: Parola C, Salogni L, Vaira X, Scutera S, Somma P, Salvi V, Musso T, Tabbia G, Bardessono M, Pasquali C, Mantovani A, Sozzani S, Bosisio D
    PLoS ONE, 2013;8(12):e82867.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Plasma levels of procalcitonin and eight additional inflammatory molecules in febrile neutropenic patients.
    Authors: Neuenschwander LC, Bittencourt H, Ribeiro AF, Teixeira AL, Teixeira MM, Teixeira JC, Nobre V
    Clinics (Sao Paulo), 2011;66(10):1699-705.
    Species: Human
    Sample Types: Plasma
  16. Abnormalities in serum chemokine levels in euthymic patients with bipolar disorder.
    Authors: Brietzke E, Kauer-Sant'Anna M, Teixeira AL, Kapczinski F
    Brain Behav. Immun., 2009;23(8):1079-82.
    Species: Human
    Sample Types: Serum
  17. Lenalidomide inhibits osteoclastogenesis, survival factors and bone-remodeling markers in multiple myeloma.
    Authors: Breitkreutz I, Raab MS, Vallet S, Hideshima T, Raje N, Mitsiades C, Chauhan D, Okawa Y, Munshi NC, Richardson PG, Anderson KC
    Leukemia, 2008;22(10):1925-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. p38 mitogen-activated protein kinase inhibitor LY2228820 enhances bortezomib-induced cytotoxicity and inhibits osteoclastogenesis in multiple myeloma; therapeutic implications.
    Authors: Ishitsuka K, Hideshima T, Neri P, Vallet S, Shiraishi N, Okawa Y, Shen Z, Raje N, Kiziltepe T, Ocio EM, Chauhan D, Tassone P, Munshi N, Campbell RM, Dios AD, Shih C, Starling JJ, Tamura K, Anderson KC
    Br. J. Haematol., 2008;141(5):598-606.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Increased serum levels of CCL11/eotaxin in schizophrenia.
    Authors: Teixeira AL, Reis HJ, Nicolato R, Brito-Melo G, Correa H, Teixeira MM, Romano-Silva MA
    Prog. Neuropsychopharmacol. Biol. Psychiatry, 2008;32(3):710-4.
    Species: Human
    Sample Types: Serum
  20. Altered levels of CC chemokines during pulmonary CMV predict BOS and mortality post-lung transplantation.
    Authors: Weigt SS, Elashoff RM, Keane MP, Strieter RM, Gomperts BN, Xue YY, Ardehali A, Gregson AL, Kubak B, Fishbein MC, Saggar R, Ross DJ, Lynch JP, Zisman DA, Belperio JA
    Am. J. Transplant., 2008;8(7):1512-22.
    Species: Human
    Sample Types: BALF
  21. Association between the macrophage inflammatory protein-l alpha gene polymorphism and Alzheimer's disease in the Chinese population.
    Authors: Li K, Dai D, Yao L, Gu X, Luan K, Tian W, Zhao Y, Wang B
    Neurosci. Lett., 2008;433(2):125-8.
    Species: Human
    Sample Types: Plasma
  22. Clinical, radiographic, and biochemical characterization of multiple myeloma patients with osteonecrosis of the jaw.
    Authors: Raje N, Woo SB, Hande K, Yap JT, Richardson PG, Vallet S, Treister N, Hideshima T, Sheehy N, Chhetri S, Connell B, Xie W, Tai YT, Szot-Barnes A, Tian M, Schlossman RL, Weller E, Munshi NC, Van Den Abbeele AD, Anderson KC
    Clin. Cancer Res., 2008;14(8):2387-95.
    Species: Human
    Sample Types: Serum
  23. Impact of tobacco-smoke on key signaling pathways in the innate immune response in lung macrophages.
    Authors: Birrell MA, Wong S, Catley MC, Belvisi MG
    J. Cell. Physiol., 2008;214(1):27-37.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Successful HAART is associated with high B-chemokine levels in chronic HIV type 1-infected patients.
    Authors: Brito A, Almeida A, Gonsalez CR, Mendonca M, Ferreira F, Fernandes SS, Duarte AJ, Casseb J
    AIDS Res. Hum. Retroviruses, 2007;23(7):906-12.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. Novel role for the liver X nuclear receptor in the suppression of lung inflammatory responses.
    Authors: Birrell MA, Catley MC, Hardaker E, Wong S, Willson TM, McCluskie K, Leonard T, Farrow SN, Collins JL, Haj-Yahia S, Belvisi MG
    J. Biol. Chem., 2007;282(44):31882-90.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. Targeting MEK1/2 blocks osteoclast differentiation, function and cytokine secretion in multiple myeloma.
    Authors: Breitkreutz I, Raab MS, Vallet S, Hideshima T, Raje N, Chauhan D, Munshi NC, Richardson PG, Anderson KC
    Br. J. Haematol., 2007;139(1):55-63.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Role of matrix metalloproteinases in the inflammatory response in human airway cell-based assays and in rodent models of airway disease.
    Authors: Birrell MA, Wong S, Dekkak A, De Alba J, Haj-Yahia S, Belvisi MG
    J. Pharmacol. Exp. Ther., 2006;318(2):741-50.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. Modification of TLR-induced activation of human dendritic cells by type I IFN: synergistic interaction with TLR4 but not TLR3 agonists.
    Authors: Walker J, Tough DF
    Eur. J. Immunol., 2006;36(7):1827-36.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. Manipulation of host cytokine network by ticks: a potential gateway for pathogen transmission.
    Authors: Hajnicka V, Vancova I, Kocakova P, Slovak M, Gasperik J, Slavikova M, Hails RS, Labuda M, Nuttall PA
    Parasitology, 2005;130(0):333-42.
    Species: Human
    Sample Types: Saliva
  30. 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
  31. Presence of dendritic cells, MCP-1, and activated microglia/macrophages in amyotrophic lateral sclerosis spinal cord tissue.
    Authors: Henkel JS, Engelhardt JI, Siklos L, Simpson EP, Kim SH, Pan T, Goodman JC, Siddique T, Beers DR, Appel SH
    Ann. Neurol., 2004;55(2):221-35.
    Species: Human
    Sample Types: CSF
  32. Involvement of Fc gamma RI (CD64) in the mechanism of HIV-1 inhibition by polyclonal IgG purified from infected patients in cultured monocyte-derived macrophages.
    Authors: Holl V, Hemmerter S, Burrer R, Schmidt S, Bohbot A, Aubertin AM, Moog C
    J. Immunol., 2004;173(10):6274-83.
    Species: Human
    Sample Types: Cell Culture Supernates
  33. Gene copy number regulates the production of the human chemokine CCL3-L1.
    Authors: Townson JR, Barcellos LF, Nibbs RJ
    Eur. J. Immunol., 2002;32(10):3016-26.
    Species: Human
    Sample Types: Cell Culture Supernates

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Reviews for Human CCL3/MIP-1 alpha DuoSet ELISA

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Human CCL3/MIP-1 alpha DuoSet ELISA
By Anonymous on 02/11/2020
Application: Sample Tested: K562 human chronic myelogenous leukemia cell line

Human CCL3/MIP-1 alpha DuoSet ELISA
By Anonymous on 12/11/2019
Application: Sample Tested: human whole blood

Human CCL3/MIP-1 alpha DuoSet ELISA
By Anonymous on 08/02/2016
Application: Sample Tested: THP-1 human acute monocytic leukemia cell line