Multiplexed Protein Detection; Detection of Relative Protein Expression; Detecting Patterns of Cytokine Expression; Biomarker/ Key Factor Screening; Identifying Key Factors; Confirming a Biological Process
Mouse Cytokine Antibody Array C3 Membranes
Wash Buffer 1
Wash Buffer 2
Biotinylated Detection Antibody Cocktail
Detection Buffer C
Detection Buffer D
8-Well Incubation Tray
Other Materials Required
Pipettors, pipet tips and other common lab consumables
Orbital shaker or oscillating rocker
Tissue Paper, blotting paper or chromatography paper
Adhesive tape or Saran Wrap
Distilled or de-ionized water
A chemiluminescent blot documentation system (such as UVP's ChemiDoc-It® or EpiChem II Benchtop Darkroom or GE's ImageQuant™ LAS 4000 or Amersham Imagers 600 and 680), X-ray Film and a suitable film processor, or another chemiluminescent detection system.
Incubate with Sample
Incubate with Biotinylated Detection Antibody Cocktail
Incubate with HRP-Conjugated Streptavidin
Incubate with Detection Buffers
Image with chemiluminescent imaging system
Perform densitometry and analysis
For best results, store the entire kit frozen at -20°C upon arrival. Stored frozen, the kit will be stable for at least 6 months which is the duration of the product warranty period. Once thawed, store array membranes and 1X Blocking Buffer at -20°C and all other reagents undiluted at 4°C for no more than 3 months.
We used the assay to analyze lung tissue extracts at 500ug per membrane. We got a good signal and reproducible results. However, I would recommend using different ECL reagent for detection, the one provided with the kit is not the best.
Michael P. Schonfeld University of Kansas Medical Center06/17/2020, 01:57 PM
The protocol is easy and user friendly. However, the read out proved difficult as the radiance between samples in different conditions can not be discerned clearly even when using advanced western blot readout software. Tried different exposures and did not get adequate results for a manuscript figure.
Laurence National Cancer Institute, NIH12/14/2019, 11:36 AM
We used the above kit to get the cytokine patterns in the secretome of the infected bone marrow derived macrophages. The kit seems to be consistent. With in vitro stimulated cells, the cytokine levels were found. We used undiluted samples in analysis. The FBS from VWR at 2% was also present in the medium.
Dipti Singh University of Chicago10/18/2019, 09:14 PM
We used the AAM-CYT-3-8 Kit. With the detailed protocol manual, we could take all the necessary precautions to get consistent signals with the positive controls and we could confirm the same after repeating it for the selected individual cytokines that were deferentially unregulated
Damini Singh W.M. Keck Center for Transgene Research10/18/2019, 03:30 PM
This product is fantastic! We use to analyze the connection between irradiation dose and inflammation in mouse plasma in mice with different genetic backgrounds. The protocol is easy to follow and the results are always clear to interpret.
Yekaterina Leonova Roswell Park Cancer Institute06/13/2019, 10:11 AM
I used this array in the serum samples of deer mice and got good pictures. The images can be easily analyzed by Imagelab.
Owen University of South Carolina01/07/2019, 10:06 PM
Protocol is easy to follow and convenient to use. It is time-saving.
Christina Purdue University05/23/2018, 05:48 PM
I used this array to analyze some astrocyte culture-conditioned (serum-free) media (about 1 million cells/well). I didn't use the software for analysis, and it took me about a day to analyze the four membranes. I didn't dilute the media and I certainly wasn't maxing out the signal when going with all of the optional overnight incubations.