The largest database of trusted experimental protocols

Proteome profiler mouse cytokine array panel a kit

Manufactured by R&D Systems
Sourced in United States

The Proteome Profiler Mouse Cytokine Array Panel A Kit is a tool for the simultaneous detection and quantification of 40 different mouse cytokines, chemokines, and other soluble proteins in a single experiment. It is designed to provide a broad overview of the relative levels of multiple analytes in a sample. The kit includes a membrane-based antibody array, buffers, and other necessary components for sample preparation and incubation.

Automatically generated - may contain errors

19 protocols using proteome profiler mouse cytokine array panel a kit

1

Profiling Cytokines in ICH Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
The inflammatory cytokine/chemokine levels in the serum of sham mice and ICH mice on day 3 after ICH were analyzed using the Mouse Cytokine Proteome Profiler Array Panel A kit (R&D system, Minneapolis, MN, USA) according to the manufacturer’s instructions. Briefly, 200 µL serum samples mixed with array buffers and detection antibody cocktails were incubated overnight at 4 °C on a platform shaker with pre-coated captured antibody array membranes. The signal was developed with West-Femto Maximum Sensitivity Substrate (Thermofisher Scientific), and images were captured with a Syngene Imaging System (Frederick, MD, USA). ImageJ-protein array was used to quantify spot density; then, the background signal was subtracted from each value. Finally, we compared the spot density on different arrays to evaluate the relative changes in cytokine/chemokine levels between sham and day 3 ICH groups. A total of 8 mice were used in this experiment (n = 4 sham and n = 4 ICH).
+ Open protocol
+ Expand
2

Quantifying Brain Cytokine Profiles

Check if the same lab product or an alternative is used in the 5 most similar protocols
The inflammatory cytokines in the mice brain were analyzed using the Mouse Cytokine Proteome Profiler Array Panel A kit (R&D Systems) according to the manufacturer’s specifications. Briefly, 300 μg mice brain samples were incubated for 1 h at room temperature with the supplied cytokine array panel A antibody cocktail. The array membranes were blocked with the blocking buffer, followed by incubating with the lysate-antibody mixtures overnight at 4 ℃ on the platform shaker. After washing in wash buffer, the array membranes were further incubated with streptavidin-HPR in blocking buffer for 30 min at room temperature before mixing with the Chemi reagent mix. Images were then captured with Bio-Rad ChemiDoc™ MP Imaging System. ImageJ was used to quantify and determine spot density.
+ Open protocol
+ Expand
3

Serum Cytokine Profiling in TNFR1/R2-Deficient Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
Analysis of serum cytokines and chemokines at multiple time points was done in male mice to identify both increased and decreased levels of several pro- and anti-inflammatory cytokines in TNFR1/R2−/− for comparison to wild type mice. On Days 1, 14, and Week 18 (n = 3/group), groups of mice were deeply anesthetized, and blood was collected transcardially into microtainer tubes (BD Diagnostics, Franklin Lakes, NJ, USA). Samples were kept on ice for 60 min to allow clotting, centrifuged at 5000 × g for 5 min, and the serum stored at −80°C for analysis. The Proteome Profiler mouse cytokine array Panel A Kit (#ARY006, R&D Systems, Minneapolis, MN, USA) was used to quantify 40 different cyto- and chemokines simultaneously by membrane-based dot blot following manufacturer’s instructions. Digitalized radiographic films were analyzed using NIH ImageJ software and values expressed as relative pixel density.
+ Open protocol
+ Expand
4

Mouse Cytokine Array Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
This assay was accomplished with Proteome Profiler Mouse Cytokine Array Panel A kit (Cat#ARY006, R&D system)35 (link). For each membrane, 200 µl of protein solution was mixed with 100 µl of sample buffer (array buffer 4) and 1.2 ml of block buffer (array buffer 6), then added with 15 µl of reconstituted Mouse Cytokine Array Panel A Detection Antibody Cocktail and incubated at room temperature for 1 h. The array membrane was incubated with block buffer (array buffer 6) for 2 h on a rocking platform shaker in the meantime, and then the block buffer was aspirated, the prepared sample/antibody mixture was added onto the membrane and incubated overnight at 4 °C on a rocking platform shaker. The membrane was washed with 20 ml of 1× wash buffer for 10 min on a rocking platform shaker for three time and rinsed with deionized water once, then was probed with Fluorophore-conjugated streptavidin (1:5,000 dilution, Cat#926-32230, Li-Cor) at room temperature for 30 min on a rocking platform shaker, washed with wash buffer for three times and rinsed with deionized water once again as in above steps. Antibody-antigen complexes were visualized using Odyssey Detection (Li-Cor, Serial No. ODY-2329) at 800 nm wavelengths. The densities of the spots were analyzed by Image J software.
+ Open protocol
+ Expand
5

Mouse Cytokine Array Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
This assay was accomplished with Proteome Profiler Mouse Cytokine Array Panel A kit (Cat#ARY006, R&D system)35 (link). For each membrane, 200 µl of protein solution was mixed with 100 µl of sample buffer (array buffer 4) and 1.2 ml of block buffer (array buffer 6), then added with 15 µl of reconstituted Mouse Cytokine Array Panel A Detection Antibody Cocktail and incubated at room temperature for 1 h. The array membrane was incubated with block buffer (array buffer 6) for 2 h on a rocking platform shaker in the meantime, and then the block buffer was aspirated, the prepared sample/antibody mixture was added onto the membrane and incubated overnight at 4 °C on a rocking platform shaker. The membrane was washed with 20 ml of 1× wash buffer for 10 min on a rocking platform shaker for three time and rinsed with deionized water once, then was probed with Fluorophore-conjugated streptavidin (1:5,000 dilution, Cat#926-32230, Li-Cor) at room temperature for 30 min on a rocking platform shaker, washed with wash buffer for three times and rinsed with deionized water once again as in above steps. Antibody-antigen complexes were visualized using Odyssey Detection (Li-Cor, Serial No. ODY-2329) at 800 nm wavelengths. The densities of the spots were analyzed by Image J software.
+ Open protocol
+ Expand
6

Cytokine Profiling of C. parapsilosis Infection

Check if the same lab product or an alternative is used in the 5 most similar protocols
BMDMs were infected with C. parapsilosis (GA1 strain; MOI of 5:1) for 24 h in 24-well plates, and cell culture supernatants were pooled from at least 3 independent experiments. The Proteome Profiler mouse cytokine array panel A kit (R&D Systems) was applied for multiplex detection of cytokines according to the manufacturer’s instructions (https://resources.rndsystems.com/pdfs/datasheets/ary006.pdf). A total of 700 μl pooled supernatant was used for a single membrane. Chemiluminescence was visualized by Image Studio Digits 3.1.
+ Open protocol
+ Expand
7

Cytokine Array of CAR T-cell Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
Sera from CAR WT and CARHEPKO mice were used for cytokine array. The array was performed using Proteome Profiler Mouse Cytokine Array Panel A Kit (R&D systems #ARY006) according to the manufacturer’s instruction. Briefly, membranes were blocked for one hour at room temperature. Meanwhile, samples were incubated with detection antibody for one hour at room temperature. Then, the blocked membranes were incubated in the sample/detection antibody mixtures overnight at 4 °C. Next day, the membranes were washed three times and incubated with diluted Streptavidin-HRP for 30 min. at room temperature. After washing three times, the membranes were incubated with Chemi Reagent Mix and each spot was visualized on ChemiDoc™ touch imaging system (Bio-Rad). The signal intensities of each spot were calculated using ImageJ software, and each value was normalized against the mean values of reference spots (pre-defined by the manufacturer).
+ Open protocol
+ Expand
8

Cytokine Profiling in Mouse Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
ELISA kits for mouse IL-6, TNFα, IL-1β, CCL2, CXCL1, CCL5, CCL3, CCL4, and CXCL2, and a Proteome Profiler Mouse Cytokine Array Panel A kit were purchased from R&D Systems and used to measure cytokines in culture media and blood serum according to the manufacturer’s instructions. Synergy HTX (Agilent Technologies) reader was used for the measurement of absorbance with Gen5 software (version 2.04) for data collection and analyses.
+ Open protocol
+ Expand
9

Serum Cytokine Profiling Using Mouse Array

Check if the same lab product or an alternative is used in the 5 most similar protocols
The Proteome Profiler Mouse Cytokine Array Panel A kit (ARY006; R&D Systems, Minneapolis, MN, USA) was used to monitor serum inflammatory cytokine profiles, which was performed according to the manufacturer’s instructions. Equal volumes (200 μL) of serum and assay buffer (a cocktail of biotinylated detection antibodies) were mixed and then incubated with cytokine pre-coated membranes. The membranes were incubated with HRP-conjugated secondary antibody cocktail, and then the cytokines were detected as dot blots by ChemiDoc™ Imaging Systems (Bio-Rad, Hercules, CA, USA). The dot blots were quantified using ImageJ software (National Institutes of Health, Bethesda, MD, USA).
+ Open protocol
+ Expand
10

Cytokine and Chemokine Profiling in Lung

Check if the same lab product or an alternative is used in the 5 most similar protocols
To examine cytokine and chemokine expression in lung samples, a total of 200 µg protein was analyzed using the Proteome Profiler Mouse Cytokine Array Panel A Kit (ARY006, R&D Systems, USA). The expression of cytokines and chemokines was determined densitometrically using Fiji software after subtracting the negative control’s average signal. Values were expressed as % of T/SS [35 (link)].
+ 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!