Human CCL4/MIP-1 beta DuoSet ELISA

Catalog # Availability Size / Price Qty
DY271
DY271-05
Ancillary Products Available
Human CCL4 / MIP-1 beta ELISA Standard Curve
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Product Details
Procedure
Citations (18)
FAQs
Supplemental Products
Reviews (3)

Human 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 human 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

Human 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 Human 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.

18 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.
    Species: Human
    Sample Types: Serum
  2. 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
  3. New Insights into the Pro-Inflammatory Activities of Ang1 on Neutrophils: Induction of MIP-1? Synthesis and Release
    PLoS ONE, 2016;11(9):e0163140.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
    Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
    PLoS ONE, 2015;10(8):e0134619.
    Species: Human
    Sample Types: Plasma
  5. 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
  6. Human intestinal epithelial cells respond to beta-glucans via Dectin-1 and Syk.
    Authors: Cohen-Kedar S, Baram L, Elad H, Brazowski E, Guzner-Gur H, Dotan I
    Eur J Immunol, 2014;44(12):3729-40.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. 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
  8. 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
  9. A novel p38-MAPK signaling axis modulates neutrophil biology in head and neck cancer.
    Authors: Dumitru CA, Fechner MK, Hoffmann TK
    J. Leukoc. Biol., 2012;91(4):591-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. CCL4L Polymorphisms and CCL4/CCL4L Serum Levels Are Associated with Psoriasis Severity.
    Authors: Pedrosa E, Carretero-Iglesia L, Boada A, Colobran R, Faner R, Pujol-Autonell I, Palou E, Esteve A, Pujol-Borrell R, Ferrandiz C, Juan M, Carrascosa JM
    J. Invest. Dermatol., 2011;131(9):1830-7.
    Species: Human
    Sample Types: Serum
  11. 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
  12. Fungal proteases induce Th2 polarization through limited dendritic cell maturation and reduced production of IL-12.
    Authors: Lamhamedi-Cherradi SE, Martin RE, Ito T, Kheradmand F, Corry DB, Liu YJ, Moyle M
    J. Immunol., 2008;180(9):6000-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Human anti-inflammatory macrophages induce Foxp3+ GITR+ CD25+ regulatory T cells, which suppress via membrane-bound TGFbeta-1.
    Authors: Savage ND, De Boer T, Walburg KV, Joosten SA, van Meijgaarden K, Geluk A, Ottenhoff TH
    J. Immunol., 2008;181(3):2220-6.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. 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
  15. Human epithelial cells establish direct antifungal defense through TLR4-mediated signaling.
    Authors: Weindl G, Naglik JR, Kaesler S, Biedermann T, Hube B, Korting HC, Schaller M
    J. Clin. Invest., 2007;117(12):3664-3672.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. 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
  17. 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
  18. High avidity antigen-specific CTL identified by CD8-independent tetramer staining.
    Authors: Choi EM, Chen JL, Wooldridge L, Salio M, Lissina A, Lissin N, Hermans IF, Silk JD, Mirza F, Palmowski MJ, Dunbar PR, Jakobsen BK, Sewell AK, Cerundolo V
    J. Immunol., 2003;171(10):5116-23.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

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Reviews for Human CCL4/MIP-1 beta DuoSet ELISA

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Human CCL4/MIP-1 beta DuoSet ELISA
By Anonymous on 02/17/2020
Application: Sample Tested: K562 human chronic myelogenous leukemia cell line

Human CCL4/MIP-1 beta DuoSet ELISA
By Anonymous on 01/03/2019
Application: Sample Tested: Cell culture supernatant

Human CCL4/MIP-1 beta DuoSet ELISA
By Anonymous on 02/27/2018
Application: Sample Tested: human whole blood