At the end of the experiment, mouse serum was harvested and stored at −80°C for Luminex multiplex analysis. Multiplex analysis was performed by personnel at Woongbee Meditech Biotechnology Inc. (Seoul, Korea), and serum levels of mouse insulin-like growth factor-1, insulin-like growth factor-binding protein (IGFBP)-1, IGFBP-3, and human IGFBP-1 were determined. Standard curves for each cytokine kit (LXSAMSM-01, LXSAMSM-02, and LXSAHM-01; R&D Systems, Minneapolis, MN, USA) were generated using the reference cytokine concentrations provided by the manufacturer.
Lxsahm 01
Manufactured by R&D Systems
Sourced in United States
The LXSAHM-01 is a laboratory equipment product. It is designed for specialized scientific applications. The core function of this product is to perform specific tasks within a controlled laboratory environment. However, a detailed description while maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
4 protocols using lxsahm 01
1
Serum Protein Analysis in Mice
2
Customized Multiplex Cytokine Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In this study, we used a custom-designed Human Luminex Discovery Assay LXSAHM-20 (CCL2, CCL3, CCL4, CD25/sIL-2Rα, CXCL9, CXCL10, IFN-γ, IFN-γ R1, IL-1α, IL-1β, IL-2, IL-6, IL-8, IL-10, IL-12, IL-17, IL-21, IL-33, TNF-α, VEGF) and LXSAHM-01 (separately CCL5 and IL-23) (R&D systems, McKinley, MN, USA) with some modifications according to the manufacturer’s instructions. The following adjustments were made: i) one additional standard was included in the three-fold serial dilution making the standard range from 1:3 to 1:2.187; (ii) samples were diluted 1:2 for the 20-plex and IL-23, and 1:50 for CCL5. Data was acquired on a Luminex 100 System (Luminex Corp., Austin, TX, USA) and StarStation Software v.3.0 [Applied Cytometry System, Dinnington, UK). The five-parameter logistic algorithm (weighted by 1/y, (V2.4)] and raw median fluorescence intensity values were used for the creation of standard curves.
3
Quantification of Fibrillin-1, Vitamin D-Binding Protein, and SERPINF1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
4
Multiplex Biomarker Measurement Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The selected biomarkers were all measured with a bead-based immunoassay (Luminex) in EDTA-plasma. Protein expression levels were measured on a Luminex FLEXMAP 3D, according to the manufacturer's protocol, with a minor adjustment. All reagent volumes were halved to save sample material and samples were measured in duplicate on the same plate.
The biomarkers TNF related apoptosis inducing ligand (TRAIL), human interferon-inducible protein 10 (IP-10), interferon gamma (IFNG) and Interleukin 4 (IL-4), Interleukin-6 (IL-6) and PCT were measured with one kit (LXSAHM-06, R&D systems), while Lipocalin 2 (LCN2) was measured in a separate kit (LXSAHM-01, R&D systems). Raw sample data was analyzed using Bio-plex Manager (Bio-rad). Calibration curves were in duplicate and backcalculated using a 5-parameters logistic regression analysis and outlier analysis of 20% coefficient of variation was applied to both the calibrator and the samples. Values that were out of range, were replaced by the closest calibrator point and marked as out of range.
The biomarkers TNF related apoptosis inducing ligand (TRAIL), human interferon-inducible protein 10 (IP-10), interferon gamma (IFNG) and Interleukin 4 (IL-4), Interleukin-6 (IL-6) and PCT were measured with one kit (LXSAHM-06, R&D systems), while Lipocalin 2 (LCN2) was measured in a separate kit (LXSAHM-01, R&D systems). Raw sample data was analyzed using Bio-plex Manager (Bio-rad). Calibration curves were in duplicate and backcalculated using a 5-parameters logistic regression analysis and outlier analysis of 20% coefficient of variation was applied to both the calibrator and the samples. Values that were out of range, were replaced by the closest calibrator point and marked as out of range.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!