CCL5 ELISA Kit

Produktnummer ABIN625174

Produktdetails CCL5 ELISA Kit

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

Reaktivität: Maus

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 1-200 pg/mL

Untere Nachweisgrenze: 1 pg/mL

Applikation: ELISA

Verwendungszweck: Mouse RANTES (CCL5) ELISA Kit for cell culture supernatants, plasma, and serum samples.

Proben: Plasma, Cell Culture Supernatant, Serum

Analytische Methode: Quantitative

Spezifität: This ELISA kit shows no cross-reactivity with the following cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN- gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP- 5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, PSelectin, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.

Sensitivität: 1 pg/mL

Produktmerkmale:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Bestandteile:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Benötigtes Material:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Anwendungsinformationen

Applikationshinweise: Recommended Dilution for serum and plasma samples3 - 30 fold

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Aufbereitung der Reagenzien:

1. Bring all reagents and samples to room temperature (18 - 25°C) before use. 2. Sample dilution: Assay Diluent A (Item D) is used for dilution of serum/plasma samples, Assay Diluent C (Item L) can be used for dilution of cell culture supernates. 3. Assay Diluent B should be diluted 5-fold with deionized or distilled water before use. 4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µl Assay Diluent A (for serum/plasma samples) or Assay Diluent C (for cell culture supernates) into Item C vial to prepare a 50 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 4 µl RANTES standard (50 ng/ml) from the vial of Item C, into a tube with 996 µl Assay Diluent A or Assay Diluent C to prepare a 200 pg/ml standard solution. Pipette 300 µl Assay Diluent A or Assay Diluent C into each tube. Use the 200 pg/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Gently vortex to mix. Assay Diluent A or Assay Diluent C serves as the zero standard (0 pg/ml). 5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer. 6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µl of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4°C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure. 7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 10,000-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 2 µl of HRP-Streptavidin concentrate into a tube with 198 µl 1x Assay Diluent B to prepare a 100-fold diluted HRP- Streptavidin solution (do not store the diluted solution for next day use). Mix through and then pipette 100 µl of prepared 100-fold diluted solution into a tube with 10 ml 1x Assay Diluent B to prepare a final 10,000 fold diluted HRP-Streptavidin solution.

Testdurchführung:

1. Bring all reagents and samples to room temperature (18 - 25°C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µl of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4°C with gentle shaking. 3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 µl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. 4. Add 100 µl of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking. 5. Discard the solution. Repeat the wash as in step 3. 6. Add 100 µl of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking. 7. Discard the solution. Repeat the wash as in step 3. 8. Add 100 µl of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking. 9. Add 50 µl of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Ergebnisberechnung:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: -20 °C

Informationen zur Lagerung: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Haltbarkeit: 6 months

Referenzen für CCL5 ELISA Kit (ABIN625174)

Tilgner, von Trotha, Gombert, Jacobs, Drechsler, Döring, Soehnlein, Grommes: "Aspirin, but Not Tirofiban Displays Protective Effects in Endotoxin Induced Lung Injury." in: PLoS ONE, Vol. 11, Issue 9, pp. e0161218, (2016) (PubMed).

Bruce, Ilagan, Guthrie, Rivera, Choudhury, Sangha, Spencer, Bertram, Jain, Kelley, Basu: "Selected renal cells modulate disease progression in rodent models of chronic kidney disease via NF-κB and TGF-β1 pathways." in: Regenerative medicine, Vol. 10, Issue 7, pp. 815-39, (2016) (PubMed).

Escudero, Martinez de Marañón, Collado, Gonzalez-Navarro, Hermenegildo, Peiró, Piqueras, Sanz et al.: "Combined sub-optimal doses of rosuvastatin and bexarotene impair angiotensin II-induced arterial mononuclear cell adhesion through inhibition of Nox5 signaling pathways and increased RXR/PPAR? and ..." in: Antioxidants & redox signaling, Vol. 22, Issue 11, pp. 901-20, (2015) (PubMed).

Koppe, Högner, Doehn, Müller, Witzenrath, Gutbier, Bauer, Pribyl, Hammerschmidt, Lohmeyer, Suttorp, Herold, Opitz et al.: "Streptococcus pneumoniae stimulates a STING- and IFN regulatory factor 3-dependent type I IFN production in macrophages, which regulates RANTES production in macrophages, cocultured alveolar ..." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 188, Issue 2, pp. 811-7, (2012) (PubMed).

Ko, Linfert, Jang, Higbee, Watkins, Cheadle, Liu, Racusen, Grigoryev, Rabb: "Transcriptional analysis of infiltrating T cells in kidney ischemia-reperfusion injury reveals a pathophysiological role for CCR5." in: American journal of physiology. Renal physiology, Vol. 302, Issue 6, pp. F762-73, (2012) (PubMed).

Details zu CCL5

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

Andere Bezeichnung: RANTES / CCL5 (CCL5 Produkte)

Synonyme: Rantes ELISA Kit, Scya5 ELISA Kit, D17S136E ELISA Kit, RANTES ELISA Kit, SCYA5 ELISA Kit, SIS-delta ELISA Kit, SISd ELISA Kit, TCP228 ELISA Kit, eoCP ELISA Kit, MuRantes ELISA Kit, C-C motif chemokine 5-like ELISA Kit, C-C motif chemokine ligand 5 ELISA Kit, chemokine (C-C motif) ligand 5 ELISA Kit, LOC100230935 ELISA Kit, CCL5 ELISA Kit, Ccl5 ELISA Kit

Hintergrund: C-C motif chemokine 5 (MuRantes) (SIS-delta) (Small-inducible cytokine A5) (T-cell-specific protein RANTES)

Gen-ID: 20304

UniProt: P30882

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