The largest database of trusted experimental protocols

Mouse cytokine array panel a kit

Manufactured by R&D Systems
Sourced in United States, Italy

The Mouse Cytokine Array Panel A kit is a multiplex assay designed to detect the relative levels of 40 mouse cytokines, chemokines, and other soluble proteins in a single experiment. This kit utilizes a membrane-based format and provides a sensitive and semi-quantitative method for analyzing protein expression profiles in mouse samples.

Automatically generated - may contain errors

25 protocols using mouse cytokine array panel a kit

1

Multiplex Cytokine Analysis of DJ-1 KO Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
Multiplex cytokine arrays were used to compare serum levels of IL-1β and other cytokines in WT (n = 4) and DJ-1 KO mice (n = 4). All serum samples were analyzed by using mouse Cytokine Array Panel A kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions. Briefly, 100 (l serum from a mouse was loaded onto the surface of an array to incubate with the primary antibodies. After washing away unbound proteins, HRP-conjugated secondary antibodies were then applied to the array surface to form sandwich antigen-antibody complexes. After final wash to remove unbound antibodies, protein spots on the array surface were visualized by incubation with ECL substrate. The array images were captured by using a chemiluminescence imaging and documentation systems (UVP, Upland, CA, USA), and abundance of cytokines of both groups in the array images was quantified by using ImageQuant 5.0 software, and normalized against the corresponding cytokines’ levels in the WT group.
+ Open protocol
+ Expand
2

Cytokine and Chemokine Profiling in HSV-1 Infected Corneas

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cytokine and chemokine levels in HSV-1 infected corneal tissues from B6 and NK1R−/− mice were measured using a Mouse Cytokine Array Panel A kit (R&D systems, Minneapolis, MN). Corneas dissected from naïve or infected eyes (day 4 and day 9 post-infection) of B6 and NK1R−/− mice were transferred to 1x PBS with protease inhibitor cocktail (PIC) and kept at −80°C. Sonication of the corneas was performed at 50% amplitude with a cycle of 15 second pulse, followed by 1 minute of resting on ice. A total of six cycles were given to each cornea. The tissue lysates (sonicated samples) obtained were centrifuged at 4°C at 15,000 rpm for 10 minutes. The supernatant was collected and the amount of protein in each sample was estimated using a BCA protein assay kit (Thermo Scientific). A total of four corneas from each group was pooled to obtain a minimum of 200 μg of protein for cytokine and chemokine array analysis as per the manufacturer instructions. Positive signals on the membranes were quantified using Image Studio Lite Version 4.0.
+ Open protocol
+ Expand
3

Cytokine Profile Analysis of Laser-Irradiated BMSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cytokine/chemokine profiles in supernatants of the cultured BMSCs population in the laser irradiated group or untreated group (control) were assessed at the end of each set of irradiation (Day 0, 5 days, 10 days, 15 days) by using Mouse Cytokine Array Panel A kit (R&D Systems, Milano, Italy) according to the manufacturer's instructions as previously described (Sabbieti et al., 2015 (link)).
+ Open protocol
+ Expand
4

Western Blot and Cytokine Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Briefly, cells were washed with cold PBS and lysed in a buffer supplemented with protease and phosphatase inhibitors. Membranes with transferred proteins were incubated with the primary antibody anti-pAKT (Ser473) (1:1000, Cell Signaling) or anti-vinculin (1:10000). The primary antibody incubation was followed by incubation with peroxidase conjugated to the secondary antibody (anti-rat, 1:10000). A chemiluminescence reaction using the ECL Plus kit (GE Healthcare) was detected using G:BOX iChemi system (Syngene). Tumor relative levels of cytokines and chemokines were measured using the Mouse Cytokine Array Panel A kit (R&D Systems) following the manufacturer's instructions. Images of the blots were acquired with G:BOX Chemi system (Syngene) and quantitative analyses were performed using ImageJ software. The 40 cytokines and chemokines of interest were normalized to the corresponding positive controls.
+ Open protocol
+ Expand
5

Cytokine Profile Analysis in BMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cytokine/chemokine profiles in supernatants of 3-day cultured BMCs population were assessed by using Mouse Cytokine Array Panel A kit (R&D Systems, Milano, Italy) accordingly to the manufacturer’s instructions. Briefly, after 3 days in culture BMCs supernatant (600 μl) from each experimental group was incubated overnight on nitrocellulose membranes spotted with specific antibodies. Chemiluminescence detection produced signals (blots) directly proportional to the amount of cytokine bound. Immunoreactive dots were visualized using LiteAblot Turbo luminol reagents (EuroClone, Milano, Italy) and Hyperfilm-ECL film (EuroClone, Milano, Italy) and quantitated densitometrically using Image J software.
+ Open protocol
+ Expand
6

Cytokine Expression Profiling in SH005S7-Stimulated Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were stimulated with SH005S7 at the indicated concentrations for 48 hours. The cell culture medium was collected and then stored at −80°C. The supernatants were tested for cytokine expression profiles, using the mouse cytokine array panel A kit (R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer's instructions.
+ Open protocol
+ Expand
7

Cytokine/Chemokine Profiling of BMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cytokine/chemokine profiles of BMCs supernatants were assessed by ELISA-based cytokine array by using Mouse Cytokine Array Panel A kit (R&D Systems, Milano, Italy) accordingly to the manufacturer's instructions. Immunoreactive dots were visualized using LiteAblot Turbo luminol reagents (Euroclone, Milano, Italy) and Hyperfilm-ECL film (Euroclone, Milano, Italy) and quantitated densitometrically.
+ Open protocol
+ Expand
8

Cytokine Profiling in Parkinson's Model

Check if the same lab product or an alternative is used in the 5 most similar protocols
Profiling of cytokines and chemokines was conducted using a multiplex cytokine array. Briefly, protein extracted from the substantia nigra of WT and DJ-1 KO mice was quantified by using a BCA protein assay kit (Pierce, Rockford, IL), followed by analysis with a mouse Cytokine Array Panel A kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction. Proteins (200 μg / array) were loaded onto the surface of the arrays for binding the primary antibodies coated on the array surface, washed to remove the unbound proteins, and incubated with HRP-conjugated secondary antibodies to form sandwich complexes. After that, the array membranes were washed again and the arrays were developed using an enhanced chemiluminescence (ECL) substrate. The array images were then captured by a charge-coupled device (CCD) imaging and documentation system (UVP, Upland,. CA, USA). Levels of protein spots in both WT and KO groups were measured by using ImageQuant 5.0 software, and normalized against the corresponding protein spots in the WT group.
+ Open protocol
+ Expand
9

Cytokine profiling of LPS-treated cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell culture media were harvested 24h after LPS treatment from the bottom wells. One ml of each cell culture medium supernatant was collected by centrifugation at 500 x g for 5 minutes to remove particulates. Cytokines in the culture supernatant were analyzed by Mouse Cytokine Array Panel A kit (Cat# ARY006, R&D Systems, Inc. Minneapolis, MN) by following the instructions provided by the manufacturer. This array contains 40 different capture antibodies printed in duplicate to simultaneously detect differences in relative levels of cytokines and chemokines between samples. Positive signals detected by chemiluminescent reagents in the kit were exposed to the X-ray film. The pixel densities of the signals were determined by densitometric scanning using an HP Scanjet G3010 Photo Scanner and Image J V1.40 (NIH, MD). The average pixel density of the pair of duplicate spots representing each cytokine expression levels was determined by subtracting an average background signal (negative control spots as a background value) from each spot.
+ Open protocol
+ Expand
10

Cytokine Profiling of Pancreatic Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols
Secreted cytokines were measured using a mouse cytokine array panel A kit (R&D systems) and following the manufacturer’s protocol. Conditioned media of pancreatic cancer cell lines were collected as supernatants. The detection antibody cocktail was added to the supernatants for 1 h followed by incubation with membranes overnight at 4 °C. After washing, HRP-conjugated Streptavidin was added, and membranes were exposed with Chemi Reagent. Images were taken with an Amersham™ Imager 600 (GE Healthcare).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!