Mouse CCL5/RANTES DuoSet ELISA

R&D Systems | Catalog # DY478

R&D Systems
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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

31.2-2000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Mouse

Mouse CCL5/RANTES DuoSet ELISA 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
View all CCL5/RANTES ELISA Kits »

Product Summary for Mouse CCL5/RANTES DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse CCL5/RANTES. 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 Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Mouse CCL5/RANTES DuoSet ELISA

Mouse CCL5 / RANTES ELISA Standard Curve

Mouse CCL5 / RANTES ELISA Standard Curve

Kit Contents for Mouse CCL5/RANTES DuoSet ELISA

  • 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)

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: CCL5/RANTES

RANTES (Regulated upon Activation, Normal T cell-expressed, and presumably Secreted), also known as CCL5, a member of the CC chemokine family of inflammatory and immunoregulatory cytokines, initially discovered by subtractive hybridization as a T cell-specific molecule (1, 2). Mouse RANTES cDNA encodes a 91 amino acid (aa) residue precursor protein with a presumed signal peptide of 23 aa residues that is cleaved to generate the 68 aa residue mature protein (3). At the amino acid sequence level, mouse RANTES is 84% identical to human RANTES. Both human and mouse RANTES exhibit cross species activity. Naturally occurring human RANTES has been found to be a mixture of the 68 aa residue mature protein and a 66 aa residue amino-terminally truncated form (4, 5). Cells known to express RANTES include keratinocytes, eosinophils, platelets, endothelium, fibroblasts, respiratory epithelium, astrocytes, vascular smooth muscle, and CD4+ and CD8+ T cells (6-14). As suggested by its acronym, RANTES production by the various cell types is induced in response to cytokine stimulation (8-13). 
Like other CC chemokines, RANTES is a monocyte chemoattractant (2). RANTES can also chemoattract unstimulated CD4+/CD45RO+ memory T cells and stimulated CD4+ and CD8+ T cells with the naive and memory phenotypes. In addition, RANTES can chemoattract and degranulate eosinophils, as well as chemoattract and induce histamine release from basophils. Human RANTES has been shown to be an inhibitor of HIV infection of human mononuclear cells (15, 16). Several CC chemokine receptors, including CCR-1, CCR-3, CCR-4, and CCR-5, have been shown to bind RANTES and subsequently to transduce a signal by increasing the intracellular calcium ion level (2, 17-21).

Alternate Names

RANTES, SISd

Entrez Gene IDs

6352 (Human); 20304 (Mouse); 403522 (Canine); 493689 (Feline)

Gene Symbol

CCL5

Additional CCL5/RANTES Products

Product Documents for Mouse CCL5/RANTES DuoSet ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Mouse CCL5/RANTES DuoSet ELISA

For research use only

Citations for Mouse CCL5/RANTES DuoSet ELISA

Customer Reviews for Mouse CCL5/RANTES DuoSet ELISA (12)

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Showing  1 - 5 of 12 reviews Showing All
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  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Anonymous
    Sample Tested: RAW 264.7 mouse monocyte/macrophage cell line and Cell Culture Supernates
    Verified Customer | Posted 11/10/2019
    Mouse CCL5/RANTES DuoSet ELISA DY478
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Anonymous
    Sample Tested: Cell Lysates
    Verified Customer | Posted 08/10/2019
    Mouse CCL5/RANTES DuoSet ELISA DY478
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Anonymous
    Sample Tested: peritoneal lavage fluid
    Verified Customer | Posted 04/15/2019
    Mice (HIII and LIII strains) injected i.p. with 2,4,6,10 tetramethylpentadecane (pristane).
    Mouse CCL5/RANTES DuoSet ELISA DY478
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 10/08/2018
    Mouse CCL5/RANTES DuoSet ELISA DY478
  • Name: Anonymous
    Sample Tested: Min6 mouse insulinoma whole cell lysate
    Verified Customer | Posted 05/14/2018
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Anonymous
    Sample Tested: Peritoneal resident macrophages
    Verified Customer | Posted 09/27/2017
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Goutham Pattabiraman
    Sample Tested: Serum and Cell culture supernatant
    Verified Customer | Posted 04/21/2017
    Mouse CCL5/RANTES DuoSet ELISA DY478
  • Name: Anonymous
    Verified Customer | Posted 04/21/2017
    used for ELISA, worked perfectly.
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Daniel Zhang
    Sample Tested: Lung tissue
    Verified Customer | Posted 04/19/2017
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 10/06/2016
    Mouse CCL5/RANTES DuoSet ELISA DY478
  • Mouse CCL5/RANTES DuoSet ELISA
    Name: Austin Russon
    Sample Tested: Blood Serum and Cell Culture Supernates
    Verified Customer | Posted 06/03/2016
  • Mouse CCL5/RANTES DuoSet ELISA, 15 Plate
    Name: Anonymous
    Sample Tested: Purified mouse RANTES protein
    Verified Customer | Posted 10/26/2015
    50ul of murine RANTES Capture antibody (2ug/mL) was coated per well of a 96 well plate at 4 degrees overnight. Purified RANTES standard was diluted (7 fold) and assayed to ensure the quality of the ELISA. Mouse RANTES Detector antibody was added (2h @ RT) followed by a 20 min incubation with Streptavidin-HRP. Substrate solution was added to measure RANTES concentration according to the manufactures protocol. The standard curve looked great as can be seen by calculation of the R- squared value. Highly recommend doing ELISA's using kits from R&D. <br />Buffer: 7-point standard curve
    Mouse CCL5/RANTES DuoSet ELISA DY478

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Protocols

View specific protocols for Mouse CCL5/RANTES DuoSet ELISA (DY478):

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.

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