Immune Response Research Solutions

Cytokine Markers and More
Immune and inflammatory response are elevated in most diseases. We have strong solutions for detecting these responses. These include cytokine markers and panels.

Here's a new pub referencing use of one of our Interleukin Markers. ArdoSabir and AndiSumidarti. Interleukin-6 expression on inflammed rat dental pulp tissue after capped with Trigona sp. propolis. Saudi Journal of Biological Sciences Volume 24, Issue 5, July 2017, Pages 1034-1037. doi.org/10.1016/j.sjbs.2016.12.019 ...Four rats were sacrificed at 6 h, 2 days, 4 days and 7 days respectively. The teeth and surrounding bone were resected, fixed in Bouin’s fixative for 24 h, decalcified with acetic acid/formal saline for 7 days, embedded in paraffin and sectioned serially at 6 μm thickness. The sections were stained with IL-6 monoclonal antibody (Neuromics, USA) using immunohistochemistry method and viewed by light microscopy...

Image: Interleukin-6 expression on rats dental pulp tissue. Normal dental pulp with no treatment (negative control) in groups I (A–B) and inflamed dental pulp in groups II (C–D), III (E–F), IV (G–H) and V (I–J), capped with Ethanolic Extract Propolis (EEP), Extract Flavonoid-Propolis (EFP), Extract Non-Flavonoid Propolis (ENFP), and Calcium Hydroxide (Ca(OH)2), respectively. Arrows show IL-6 expression. Immunohistochemistry method, DAB chromogen, original magnification 200×.

Proud Distributor of RayBiotech's Antibody Arrays and ELISAs

Pioneer and Market Leader

I am pleased to announce we are a distribution partner for RayBiotech. The solutions we offer include Antibody Arrays and ELISA Kits. With these additions, you now have many options for the either the qualitative, semi-quantitative or quantitative measurement of protein expression. From single antibody pair ELISAs to multiplexed arrays. The arrays enable you to simultaneously analyze up to 80 cytokine, inflammatory response, growth factor or angiogenesis related proteins.

In this posting, I focus on our new C-Series Antibody Array Kits. The kits utilize the sandwich immunoassay principle, wherein a panel of capture antibodies is printed on a nitrocellulose membrane solid support (usualy 2.5 cm x 3 cm). The array membranes are processed similarly to a Western blot (chemiluminescent readout). Signals are then visualized on x-ray film or a digital image, allowing densitometry data collection and calculation of fold-changes for each detected protein. The entire procedure can be completed in 1 day, and is simple enough to permit even the novice researcher to successfully collect data with very few pitfalls and little or no optimization.

They are proven and frequently published. You can now analyze the protein expression in you assays for a fraction of the cost of tradition methods. Here's an example: Simona Giorgini, Daniela Trisciuoglio, Chiara Gabellini, Marianna Desideri, Laura Castellini, Cristina Colarossi, Uwe Zangemeister-Wittke, Gabriella Zupi and Donatella Del Bufalo. Modulation of bcl-xL in Tumor Cells Regulates Angiogenesis through CXCL8 Expression. doi: 10.1158/1541-7786.MCR-07-0088. Mol Cancer Res August 2007 5; 7

Figure: bcl-xL overexpression in ADF glioblastoma and M14 melanoma cells increases CXCL8 expression. Image of membranes from a protein array. Membranes are probed with CM from the (A) glioblastoma control (AN8) and bcl-xL–overexpressing (AXL42 and AXL74) clones and (B) melanoma control (Mneo) and bcl-xL–overexpressing (MXL12 and MXL90) clones. In the negative control (NEG), the CM is replaced with an appropriate mock buffer according to array protocol. The intensity of protein signals for each membrane was compared with the relative positive signals by densitometric analysis. C. Schematic representation of proangiogenic factors that can be detected by the use of the membrane. The CXCL8, TGF-1β, TIMP1, TIMP2, and VEGF protein signals (two spots) are indicated by red, blue, yellow, violet, and green rectangles, respectively, in each image....The Human Angiogenesis Antibody Array I (RayBiotech. Inc.) was used according to the manufacturer's protocol to evaluate the secretion of 20 angiogenic factors into the CM of the different lines. A schematic representation of the proangiogenic factors that may be detected by the use of the array has been reported in the above figure. Membranes spotted in duplicate with antibodies against angiogenic factors were blocked with blocking buffer and then were incubated overnight with CM. Next, membranes were washed with wash buffer, incubated with biotin-conjugated antibodies against proangiogenic factors, washed with wash buffer, and incubated with horseradish peroxidase–conjugated streptavidin. The signals on the membranes were detected by chemiluminescence. Membranes, blocking and wash buffer, and antibodies against proangiogenic factors were all provided with the kit. The intensity of protein signal (two spots for each protein) was compared with the relative positive signals by densitometric analysis.

Here's an excellent video that provides more detail on Antibody Array Kit capabililities:

I will continue to post information on new addition with related pubs and data.

Quantibody® Cytokine Arrays

Cytokines, Adipokines, Growth Factors and Neurotrophins

We are pleased to announce addition of Antibody Arrays to our catalog. These are excellent tools for study apoptosis, inflammation, angiogenesis and  immune response and related autoimmune, inflammatory and degenerative diseases plus many forms of cancers. Check out this video.

Specific Research Applications Include:
•High-throughput profiling of cytokine expression
•Validation of semi-quantitative antibody array results
•Identifying potential molecular targets for drug development
•Identifying the molecular mechanisms of drug action
•Identifying crucial factors involved in disease processes
•Discovering biomarkers for disease management
•Discovering expression patterns for molecular classification of diseases

How Quantibody Arrays Work

We will be aggresively adding new arrays to our product offerings in June. Stay tuned for more.





Aging-Caloric Restriction and BDNF-Leptin

As we age, selective BDNF receptors increase in the the dorsal vagal complex (DVC), the brainstem center mediating the satiety reflex. Aging also affects the suppressor of cytokine signaling-3. This results in abnormal responses to BDNF and Leptin Signaling.

In this study, the investigators found that by restricting calories in rats, DVC BDNF immunoreactive concentrations were elevated and resulting in satiety threshold stability.  This indicates functional desensitization of the DVC to these signals: Karine Bédard, Stéphanie Segura, Stéphanie Mahaut, Catherine Tardivel, Guylaine Ferland, Bruno Lebrun, Pierrette Gaudreau. Effects of aging and caloric restriction on brainstem satiety center signals in rats. Mechanisms of Ageing and Development. dx.doi.org/10.1016/j.mad.2012.01.004.

The authors used our BDNF Antibody to determine expression in the DVC.....goat serum and 0.3% Triton-X-100) (Sigma–Aldrich), overnight at 4 °C with chicken anti-rat BDNF antibody (Neuromics, Edina, MN; diluted 1/200 in blocking buffer), rinsed 3× 10 min in fresh PBS and incubated for 2 h, in the dark...

These findings could lead to potential therapies based on modulating BDNF expression in the DVC.