Mouse CCL4/MIP-1 beta DuoSet ELISA

Catalog # Availability Size / Price Qty
DY451
DY451-05
Ancillary Products Available
Mouse CCL4 / MIP-1 beta ELISA Standard Curve
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Product Details
Procedure
Citations (15)
FAQs
Supplemental Products
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Mouse CCL4/MIP-1 beta 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 CCL4/MIP-1beta. 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 CCL4 / MIP-1 beta 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: 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); 102117861 (Cynomolgus Monkey)
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

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 CCL4/MIP-1 beta 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.

15 Citations: Showing 1 - 10
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  1. In-depth characterization of congenital Zika syndrome in immunocompetent mice: Antibody-dependent enhancement and an antiviral peptide therapy
    Authors: VN Camargos, G Foureaux, DC Medeiros, VT da Silveir, CM Queiroz-Ju, ALB Matosinhos, AFA Figueiredo, CDF Sousa, TP Moreira, VF Queiroz, ACF Dias, KTO Santana, I Passos, ALCV Real, LC Silva, FAG Mourão, NT Wnuk, MAP Oliveira, S Macari, T Silva, GP Garlet, JA Jackman, FM Soriani, MFD Moraes, EMAM Mendes, FM Ribeiro, GMJ Costa, AL Teixeira, NJ Cho, ACP Oliveira, MM Teixeira, VV Costa, DG Souza
    EBioMedicine, 2019;0(0):.
    Species: Mouse
    Sample Types: Plasma
  2. Probiotic treatment during neonatal age provides optimal protection against experimental asthma through the modulation of microbiota and T cells: Neonatal exposure to probiotics prevents asthma
    Authors: CFCG Nunes, JS Nogueira, FB Canto, PHO Vianna, BT Ciambarell, PMRE Silva, KR Miranda, LA Lobo, RMCP Domingues, M Busch, GC Atella, AM Vale, M Bellio, A Nóbrega, R Fucs
    Int. Immunol., 2018;0(0):.
    Species: Mouse
    Sample Types: BALF
    Applications: ELISA Capture
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. Infiltrating neutrophils induce allospecific CTL in response to immunization with apoptotic cells via MCP-1 production.
    Authors: Shiratsuchi Y, Iyoda T, Tanimoto N, Kegai D, Nagata K, Kobayashi Y
    J. Leukoc. Biol., 2007;81(2):412-20.
    Species: Mouse
    Sample Types: Peritoneal Lavage Fluid
  9. 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 Types: Cell Culture Supernates
  10. 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
  11. 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: Cell Culture Supernates
  12. 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
  13. 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
  14. Toll-like receptor 2 (TLR2) mediates astrocyte activation in response to the Gram-positive bacterium Staphylococcus aureus.
    Authors: Esen N, Tanga FY, DeLeo JA, Kielian T
    J. Neurochem., 2004;88(3):746-58.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  15. Orally administered CpG oligodeoxynucleotide induces production of CXC and CC chemokines in the gastric mucosa and suppresses bacterial colonization in a mouse model of Helicobacter pylori infection.
    Authors: Raghavan S, Nystrom J, Fredriksson M, Holmgren J, Harandi AM
    Infect. Immun., 2003;71(12):7014-22.
    Species: Mouse
    Sample Types: Serum

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