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Human CCL13/MCP-4 DuoSet ELISA

Catalog #: DY327
11 Citations Datasheet / COA / SDS

Discontinued Product

DY327 has been discontinued.
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Human CCL13 / MCP-4 ELISA Standard Curve
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Citations (11)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human CCL13/MCP-4 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
100 µL
Assay Range
7.8 - 500 pg/mL
Sufficient Materials
For 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 CCL13/MCP-4. 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)

 

Scientific Data

N/A Human CCL13 / MCP-4 ELISA Standard Curve View Larger

Human CCL13 / MCP-4 ELISA Standard Curve

Product Datasheets

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Product Datasheet
COA
SDS

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: CCL13/MCP-4

Human MCP-4, also known as CCL13, encodes a 98 amino acid (aa) precursor protein with a 23 aa hydrophobic signal peptide that is cleaved to yield an 8 kDa, 75 aa mature MCP-4. Mature MCP-4 lacks any potential N-glycosylation sites and shares a pyroglutamate proline motif with other human MCP proteins.

Entrez Gene IDs:
6357 (Human)
Alternate Names:
C-C motif chemokine 13; CCL13; chemokine (C-C motif) ligand 13; CKb10; MCP4; MCP-4; MCP-4Monocyte chemoattractant protein 4; MGC17134; NCC-1Monocyte chemotactic protein 4; NCC1Small-inducible cytokine A13; new CC chemokine 1; SCYA13CK-beta-10; SCYL1; small inducible cytokine subfamily A (Cys-Cys), member 13

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 CCL13/MCP-4 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.

11 Citations: Showing 1 - 10
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  1. Onchocerca volvulus-specific antibody and cellular responses in onchocerciasis patients treated annually with ivermectin for 30 years and exposed to parasite transmission in central Togo
    Authors: SI Johanns, RG Gantin, B Wangala, K Komlan, WA Halatoko, M Banla, P Karabou, AJ Luty, H Schulz-Key, C Köhler, PT Soboslay
    PloS Neglected Tropical Diseases, 2022-05-03;16(5):e0010340.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Serological profiling reveals hsa-miR-451a as a possible biomarker of anaphylaxis
    Authors: W Francuzik, K Pažur, M Dalke, S Dölle-Bier, M Babina, M Worm
    JCI Insight, 2022-04-08;0(0):.
    Species: Human
    Sample Types: Serum
  3. Echinococcus multilocularis specific antibody, systemic cytokine, and chemokine levels, as well as antigen-specific cellular responses in patients with progressive, stable, and cured alveolar echinococcosis: A 10-year follow-up
    Authors: B Grüner, L Peters, A Hillenbran, P Vo beta berg, J Schweiker, EG Rollmann, LH Rodriguez, J Blumhardt, S Burkert, P Kern, C Köhler, PT Soboslay
    PloS Neglected Tropical Diseases, 2022-02-02;16(2):e0010099.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Oxidative Stress and Inflammation Are Associated with Coexistent Severe Multivessel Coronary Artery Stenosis and Right Carotid Artery Severe Stenosis in Elderly Patients
    Authors: X Li, D Guo, Y Hu, Y Chen
    Oxidative Medicine and Cellular Longevity, 2021-12-22;2021(0):2976447.
    Species: Human
    Sample Types: Serum
  5. Vitamin D Modulates Expression of the Airway Smooth Muscle Transcriptome in Fatal Asthma.
    Authors: Himes B, Koziol-White C, Johnson M, Nikolos C, Jester W, Klanderman B, Litonjua A, Tantisira K, Truskowski K, MacDonald K, Panettieri R, Weiss S
    PLoS ONE, 2015-07-24;10(7):e0134057.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Cytokine and chemokine responses to helminth and protozoan parasites and to fungus and mite allergens in neonates, children, adults, and the elderly.
    Authors: Lechner C, Komander K, Hegewald J, Huang X, Gantin R, Soboslay P, Agossou A, Banla M, Kohler C
    Immun Ageing, 2013-07-15;10(1):29.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. IRF5 promotes inflammatory macrophage polarization and T(H)1-T(H)17 responses.
    Authors: Krausgruber T, Blazek K, Smallie T
    Nat. Immunol., 2011-01-16;12(3):231-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Genetic variants of CC chemokine genes in experimental autoimmune encephalomyelitis, multiple sclerosis and rheumatoid arthritis.
    Authors: Ockinger J, Stridh P, Beyeen AD, Lundmark F, Seddighzadeh M, Oturai A, Sorensen PS, Lorentzen AR, Celius EG, Leppa V, Koivisto K, Tienari PJ, Alfredsson L, Padyukov L, Hillert J, Kockum I, Jagodic M, Olsson T
    Genes Immun., 2009-10-29;11(2):142-54.
    Species: Human
    Sample Types: CSF
  9. CCL13 is a promising diagnostic marker for systemic sclerosis.
    Authors: Yanaba K, Yoshizaki A, Muroi E, Hara T, Ogawa F, Shimizu K, Hasegawa M, Fujimoto M, Takehara K, Sato S
    Br. J. Dermatol., 2009-09-11;162(2):332-6.
    Species: Human
    Sample Types: Serum
  10. Systemic inflammatory markers in COPD: results from the Bergen COPD Cohort Study.
    Authors: Eagan TM, Ueland T, Wagner PD, Hardie JA, Mollnes TE, Damas JK, Aukrust P, Bakke PS
    Eur. Respir. J., 2009-07-30;35(3):540-8.
    Species: Human
    Sample Types: Plasma
  11. Induction of CCL13 expression in synovial fibroblasts highlights a significant role of oncostatin M in rheumatoid arthritis.
    Authors: Hintzen C, Quaiser S, Pap T, Heinrich PC, Hermanns HM
    Arthritis Rheum., 2009-07-01;60(7):1932-43.
    Species: Human
    Sample Types: Cell Culture Supernates

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