CCL5 ELISA Kit

Catalog No. ABIN1112783

This Colorimetric ELISA kit is designed for the quantitative measurement of Human CCL5.

Quick Overview for CCL5 ELISA Kit (ABIN1112783)

Target: CCL5 (Chemokine (C-C Motif) Ligand 5 (CCL5))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.6 pg/mL - 1000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum, Tissue Lysate

Product Details CCL5 ELISA Kit

Minimum Detection Limit: 15.6 pg/mL

Purpose: For quantitative detection of Rantes in Human serum, plasma, tissue lysatesor cell culture supernatants.

Analytical Method: Quantitative

Sensitivity: < 3 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Human Rantes antibody 2. Lyophilized Human Rantes standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Human Rantes antibody (Concentrated): 130 µl.

Material not included:

1. 37 °C incubator
2. Microplate reader (wavelength: 450 nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5 mL of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1 L

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Rantes polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Rantes polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the Rantes amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of Rantes can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37 °C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Body fluids, tissue lysates and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin, EDTA or citrate as the anticoagulant. Centrifuge for 15 min at 1000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Storage: 4 °C,-20 °C

Storage Comment: Store at 2-8°C for 6 months, or at -20°C for 12 months.

Expiry Date: 12 months

Target Details for CCL5

Target: CCL5 (Chemokine (C-C Motif) Ligand 5 (CCL5))

Alternative Name: RANTES

Background: Rantes, also known as Chemokine (C-C motif) ligand 5 (also CCL5), is a protein which in humans is encoded by the CCL5 gene, this chemokine has been localized to chromosome 17 in humans. Rantes is an 8kDa protein classified as a chemotactic cytokine or chemokine. It is chemotactic for T cells, eosinophils, and basophils. The Rantes plays an active role in recruiting leukocytes into inflammatory sites. With the help of particular cytokines (i.e., IL-2 and IFN-gamma) that are released by T cells, CCL5 also induces the proliferation and activation of certain natural-killer (NK) cells to form CHAK (CC-Chemokine-activated killer) cells. It is also an HIV-suppressive factor released from CD8+ T cells. Recently, the RANTES protein has been engineered for in vivo production by Lactobacillus bacteria, and this solution is being developed into a possible HIV entry-inhibiting topical microbicide.

Pathways: Cellular Response to Molecule of Bacterial Origin, Regulation of G-Protein Coupled Receptor Protein Signaling, Smooth Muscle Cell Migration