Mouse/Rat CCL5/RANTES Quantikine ELISA Kit

R&D Systems | Catalog # MMR00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (25 µL), Platelet-poor EDTA Plasma (25 µL), Platelet-poor Citrate Plasma (25 µL)

Sensitivity

2 pg/mL

Assay Range

7.8-500 pg/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Citrate Plasma)
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Product Summary for Mouse/Rat CCL5/RANTES Quantikine ELISA Kit

The Quantikine Mouse/Rat CCL5/RANTES Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse or rat RANTES levels in cell culture supernates, serum, and platelet-poor plasma. It contains E. coli-expressed recombinant mouse RANTES as well as antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant RANTES. Results obtained using natural mouse or rat RANTES showed linear curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse and rat RANTES.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Mouse, Rat

Specificity

Natural and recombinant mouse and rat RANTES.  This assay cross-reacts 1.3% with recombinant Human RANTES.

Cross-reactivity

Cross-reactivity observed with 1 or more available related molecules. Cross-species reactivity observed with 1 or more species tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, Citrate Plasma, Platelet-poor Citrate Plasma, Platelet-poor EDTA Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 18.8 104 198 20.1 109 208
Standard Deviation 0.7 1.9 4.1 1.6 5.6 11.5
CV% 3.7 1.8 2.1 8.0 5.1 5.5

Recovery for Mouse/Rat CCL5/RANTES Quantikine ELISA Kit

The recovery of RANTES spiked to three levels in mouse samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=8) 108 98-115
Platelet-poor Citrate Plasma (n=5) 102 95-110
Platelet-poor EDTA Plasma (n=5) 99 86-110
Serum (n=5) 97 90-107

Linearity

To assess the linearity of the assay, four or more samples containing or spiked with various concentrations of RANTES in each matrix were diluted with Calibrator Diluent and then assayed.

Mouse/Rat CCL5/RANTES ELISA Linearity
Mouse/Rat CCL5/RANTES ELISA Linearity

Scientific Data Images for Mouse/Rat CCL5/RANTES Quantikine ELISA Kit

Mouse/Rat CCL5/RANTES ELISA Standard Curve

Mouse/Rat CCL5/RANTES ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. 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/Rat CCL5/RANTES Quantikine ELISA Kit

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.

Product Specific Notices for Mouse/Rat CCL5/RANTES Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Mouse/Rat CCL5/RANTES Quantikine ELISA Kit

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Protocols

View specific protocols for Mouse/Rat CCL5/RANTES Quantikine ELISA Kit (MMR00):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 50 µL Assay Diluent
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 5 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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