Snap-frozen lungs were homogenized using a Bullet Blender (Next Advance, Averill Park, NY), and total protein was isolated from lungs using protein extraction buffer as described [19 (link)]. IL-11 concentrations in lung homogenates and BAL fluid were determined using ELISA (Duoset kit, R&D systems), per the manufacturer’s instructions. G-CSF, CXCL1, CXCL2, CXCL5, CXCL10, TNFα, and IL-6 concentrations in BALF were measured using a mouse magnetic Luminex assay (R&D Systems), read using a LiquiChip 200 workstation (Qiagen, Valencia, CA).
Liquichip 200
Manufactured by Qiagen
Sourced in Canada, Switzerland
The LiquiChip 200 is a laboratory instrument designed for automated liquid handling and sample processing. It is capable of performing various liquid handling tasks such as pipetting, mixing, and plate handling. The LiquiChip 200 is intended to streamline and standardize laboratory workflows, improving efficiency and reproducibility.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using liquichip 200
1
Quantifying Lung Inflammatory Mediators
2
Quantitative Multiplexed Chemokine and Cytokine Analysis
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Chemokines and cytokines in BAL fluid supernatant were quantified using multiplexed ELISA (ProcartaPlexTM 34−plex, EBioscience). Captured analytes were detected on a Qiagen Liquichip 200 running Luminex 100 integrated system software version 2.3. Procartaplex Analyst version 1.0 was used to define standard curves and determine analyte concentrations.
3
Plasma MBD-4 Quantification in Mice
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Blood samples obtained by heart puncture of mice were centrifuged for five minutes. The supernatant was removed and stored at −20 °C until processed. Concentrations of MBD-4 in plasma samples were analysed by Luminex assay according to standard protocols using LiquiChip200 (Qiagen). A MBD-4 Elisa kit (ABIN415726, antibodies online GmbH) was used for protein detection. Every experiment was accomplished three times.
4
Multiplex Cytokine Analysis in Mice
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To analyze concentrations of different cytokines, blood samples obtained by heart puncture of the mice were centrifuged for five minutes. The supernatant was removed and stored at 20 °C until processing. The concentrations of IL-1β, IL-6, IL-10, IL-12p70 and TNFα in plasma samples were analyzed using a Luminex assay according to standard protocols with LiquiChip200 (Qiagen). A Milliplex cytokine multiplex immunoassay kit (MPXHCYTO-60 K-01; Millipore) was used for protein detection.
5
Quantification of Fibrillin-1, Vitamin D-Binding Protein, and SERPINF1
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Proteins tested were the fibrillin-1 (FBN1), Vitamin D-binding protein (VDBP), and SERPINF1. They were determined with specific assays according to manufacturer’s specifications. FBN1 was quantitated by ELISA (dilution 1:5; #MBS3804755, MyBiosource, San Diego, CA, USA), VDBP with a Multiplex assay (dilution 1:10000; #HCCBP2MAG-58K, EMD Millipore Corporation, Billerica, MA, USA), and SERPINF1, by Luminex assay (dilution 1:4000; #LXSAHM-01, R&D systems, Minneapolis, MN, USA). Protein quantification was performed using the LiquiChip 200 apparatus, and the data analysis performed with ht LiquiChip Analyzer software (Qiagen, Toronto, ON, Canada). For each biomarker, an 8-point standard curve and appropriate controls were included, and samples were done in duplicate. The minimum detectable doses were for FBN1, 0.312 ng/ml; VDBP, 0.58 ng/ml; and SERPINF1, 3.66 pg/ml.
6
Multiplex Profiling of Secreted Chemokines
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Multiplex analysis was performed on the supernatants of DC for profiling of secreted chemokines (Invitrogen, human 10-plex) according to the manufacturer’s instructions. The beads were read on a LiquiChip 200, (Qiagen, Hombrechtikon, Switzerland), and data were analyzed by the five parameter curve fitting in Luminex100 IS Software. Chemokine concentrations are presented as pg/ml. Comparison with of multiplex bead array assays with ELISA assays showed comparable results with these two quantitative analysis methods (data not shown).
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