Product nameCytokine Array – Human Cytokine Antibody Array (Membrane, 23 Targets)
See all Cytokine Antibody Array antibody arrays
Sample typeCell culture supernatant, Saliva, Milk, Urine, Serum, Plasma, Cell culture extracts, Other biological fluids, Whole Blood, Tissue Extracts, Cell Lysate, Cell culture media
Species reactivityReacts with: Human
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.
Tested applicationsSuitable for: Multiplex Protein Detectionmore details
Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 x 4 Membranes 1 x 8 Membranes 1,000X HRP-Streptavidin Buffer 1 x 50µl 1 x 50µl 1X Blocking Buffer 1 x 25ml 2 x 25ml 20X Wash Buffer I 1 x 10ml 1 x 20ml 20X Wash Buffer II 1 x 10ml 1 x 20ml 2X Cell Lysis Buffer 1 x 10ml 1 x 16ml 8-Well Incubation Tray (with Lid) 1 unit 1 unit Biotin-Conjugated Anti-Cytokines 2 vials 4 vials Cytokine Antibody Array Membranes 4 units 8 units Detection Buffer C 1 x 1.5ml 1 x 2.5ml Detection Buffer D 1 x 1.5ml 1 x 2.5ml
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.
|Multiplex Protein Detection||Use at an assay dependent concentration.|
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.