Cytokine Array – Human Cytokine Antibody Array (Membrane, 23 Targets) (ab133996)

Overview

  • Product name
    Cytokine Array – Human Cytokine Antibody Array (Membrane, 23 Targets)
    See all Cytokine Antibody Array antibody arrays
  • Sample type
    Cell culture supernatant, Saliva, Milk, Urine, Serum, Plasma, Cell culture extracts, Other biological fluids, Whole Blood, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type
    Semi-quantitative
  • Species reactivity
    Reacts with: Human
  • Product overview

    ab133996 is for simultaneous detection of 23 Human Cytokines. Suitable for all sample types.


    Targets: G-CSF, GM-CSF, GRO (alpha, beta & gamma), GRO-alpha, IL-1alpha, IL-2, IL-3, IL-5, IL-6, IL-7, IL-8, IL-10, IL-13, IL-15, IFN-gamma, MCP-1, MCP-2, MCP-3, MIG, RANTES, TGF-beta1, TNF-alpha, TNF-beta


    Cytokine arrays are an antibody-pair-based assay, analogous to ELISA, but using a membrane as a substrate rather than a plate. Capture antibodies are supplied arrayed/spotted on a membrane with each pair of spots representing a different analyte. Sample is added (0.2-1ml of 1 sample to each membrane), and then paired biotinylated detector antibodies and streptavidin HRP. The cytokine array is analyzed using the same methods as a chemiluminescent western blot. Comparison between samples can be by eye or using densitometry software for a semi-quantitative comparison.


    Learn more about membrane antibody arrays

  • Notes

    If you are interested in this cytokine array, arrays ab133997ab169804ab134003ab133998 and ab169817 may also be of interest.  


    A table listing all of our mouse membrane antibody cytokine arrays and other arrays and the analytes they measure is available here.


     


     

  • Tested applications
    Suitable for: Multiplex Protein Detectionmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab133996 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Multiplex Protein Detection Use at an assay dependent concentration.

Images

  • Human peripheral blood cells (1x106 cells/mL) were cultured in RPMI media supplemented with 10% fetal calf serum, 100 U/mL penicillin, and 100 mg/mL streptomycin sulfate.

    Cells were cultured unstimulated or stimulated with 10 mg/mL PHA. Conditioned media was harvested after 48 hours, aliquoted and assayed using ab133996. Media alone was used as a negative control.

  • Cells were cultured unstimulated or stimulated with 10 mg/mL PHA. Conditioned media was harvested after 48 hours, aliquoted and assayed using ab133996. Media alone was used as a negative control. Samples were incubated on the human cytokin antibody arrays and results were quantified using the ULTRAQuant software.

  • Abreview rating 4/5 stars. Review from Abcam user community. Verified customer. Submitted Oct 2 2014.

    The human cytokine antibody array containing 23 target proteins was used to determine the effect of lipopolysaccharide (LPS) on the induction of cytokine production in macrophages. One million macrophage cells were plated in two sets and used in the study, one set was treated with PBS alone while the other set is treated with LPS at 1 µg/mL for 24 h. After 24 h of LPS treatment, both sets of macrophages were harvested using the provided cell lysis buffer and the cells were mechanically disrupted using a 20-gauge needle. Disrupted cell lysates were centrifuged and the supernatant was collected. The protein concentration of the supernatant was determined and 200 µg of proteins diluted in the blocking buffer was applied into previously blocked membranes and stored overnight at 4 °C. The remainder of the experiment was performed using the manufacturer’s instructions where all remaining incubations were performed at room temperature for 2 h. After incubation and washing steps, the proteins were detected using the provided detection reagents and photographed in a blot imaging system. Membranes were exposed for 15 s.

Protocols

References

This product has been referenced in:
  • Keuper M  et al. Activated macrophages control human adipocyte mitochondrial bioenergetics via secreted factors. Mol Metab 6:1226-1239 (2017). Read more (PubMed: 29031722) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Q&A

Answer

Thank you for your reply. I am glad you found the information provided of help.

Each membrane of the kit should be used with a total volume of 1 mL of sample. Depending what kind of sample you are using, this may need to be pre-diluted to obtain the optimum results. A guideline of recommended sample dilutions can be found from the kit protocol descriptions:

ab133996

https://www.abcam.com/ps/products/133/ab133996/documents/ab133996%20Cytokine%20Human%20Membrane%20Antibody%20Array%2023%20Targets%20-%20protocol%20c%20array%20v2(website).pdf

ab133997

https://www.abcam.com/ps/products/133/ab133997/documents/ab133997%20Cytokine%20Human%20Membrane%20Antibody%20Array%2042%20Targets%20-%20protocol%20c%20array%20v2(website).pdf

ab133998

https://www.abcam.com/ps/products/133/ab133998/documents/ab133998%20Cytokine%20Human%20Membrane%20Antibody%20Array%2080%20Targets%20-%20protocol%20c%20array%20v2(website).pdf

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

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Answer

Thank you for contacting us and your interest in our products. My colleague xxxxx passed on your enquiry to the Scientific Support team at Abcam and I will hopefully be able to help answer your question.

The antibody arrays are a unique tool, allowing for the semi-quantitative profiling of protein expression. Each one of our kits typically allows for the simultaneous profiling of 20-80 proteins in your samples. Each kit can either be purchased with 4 or 8 membranes. From this we would typically recommend performing one positive control and one negative, allowing for 2 or 6 samples to be analysed (from the 4 or 8 membrane kits respectively). More information on these kits can be found from the following link:

www.abcam.com/arrays

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

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