Biotin Label-based Antibody Array Technology

Recent technological advances by RayBiotech have enabled the largest commercially available antibody array to date.  Biotin Label-based Antibody Arrays can be used for simultaneously detecting expression levels of 507 human or 308 mouse proteins in cell culture supernates and serum.  With the L Series Antibody Array, researchers can now obtain a broad, panoramic view of cytokine expression. 

The expression levels of 507 human or 308 mouse target proteins can be simultaneously detected, including cytokines, chemokines, adipokine, growth factors, angiogenic factors, proteases, soluble receptors, soluble adhesion molecules, and other proteins in cell culture supernate or serum. Furthermore, an internal control is used to monitor the whole process including biotin-labelling, so this massive array will accurately reflect the available cytokines in your sample. Additionally, RayBiotech have developed a new L-series Adipokine Array for specifically detecting 182 adipokine proteins in a single array.

RayBiotech offer a range of L-Series Testing Services, carried out by expert scientists at RayBiotech
Additionally, RayBiotech also offer a range of simple and easy to use L-Series Kits

How it Works

Through a simple process, your sample is biotinylated and then dialyzed in preparation for incubation with the array. The process is easy to understand and involves biotinylating the primary amine of the proteins in the sample. From here, the newly biotinylated sample is added onto the antibody arrays either membrane or glass slide and incubated at room temperature. After incubation with HRP-streptavidin or Fluorescent dye-Streptavidin, the signals can be visualized either by chemiluminescence or fluorescence.

Assay Discription

The cell culture supernates or serum is dialyzed before the biotin-labelling step. Through a simple process, the primary amine of the proteins in the sample is biotinylated, followed by dialysis to remove free biotin. From here, the newly biotinylated sample is added onto the array membrane or glass slide and incubated at room temperature. After incubation with HRP-streptavidin or Fluorescent Dye-Strepavidin, the signals can be visualized either by chemiluminescence or fluorescence.  In this array, RayBiotech use an internal control to monitor the whole process, including biotin-labelling to ensure that the array results will accurately represent the levels in your sample.  The experimental procedure is simple and can be performed in any laboratory.

L-Series Array Examples

A) Human L-Series 507

The figure shows the RayBiotech Biotin-label-based Human Antibody Array after probing with two cell culture supernates. The images were captured using a Axon GenePix laser scanner.

B) Mouse L-Series 308

The figure shows the RayBiotech Biotin-label-based Mouse Array 1 probed with Mouse sample. The image was captured using an Axon GenePix laser scanner.

Please click here for a description of other products and services offered by RayBiotech.

References

1. Profiling of cytokine expression by biotin-labeled-based protein arrays. Ying Lin, Ruochun Huang, Lipai Chen et al. Proteomics.2003, 3: 1750-1757

2. Proteomic profiling of the cancer microenvironment by antibody arrays. Vladimir Knezevic, Chidchanok Leethanakul, Verena E. Bichsel et al. Proteomics 2001, 1, 1271–1278

3. Antibody microarray profiling of human prostate cancer sera: Antibody screening and identification of potential biomarkersJeremy C. Miller, Heping Zhou, Joshua Kwekel, Robert Cavalloet al. The, Brian B. Haab. Proteomics. 2003, 3: 56-63.

4. Smad4 signalling in T cells is required for suppression of gastrointestinal cancer. Byung-Gyu Kim, Cuiling Li, WenhuiQiao, Mizuko Mamura, Barbara Kasperczak, Miriam Anver, Lawrence Wolfraim, Suntaek Hong, Elizabeth Mushinski, Michael Potter, Seong-Jin Kim, Xin-Yuan Fu, Chuxia Deng and John J. Letterio. Nature. 2006; Vol 441: 1015

5. Connexin suppresses human glioblastoma cell growth by down-regulation of monocyte chemotactic protein 1, as discoveredusing protein array technology. Ruochun Huang, Ying Lin, Cheng C. Wang, Jacob Gano, Biaoyang Lin, Qian Shi, AltonBoynton, Jocelyn Burke, and Ruo-Pan Huang. Cancer Res. 2002;62:2806-2812.

6. LPS induces the interaction of a transcription factor, LPS-induced TNF-a factor, and STAT6(B) with effects on multiple cytokines.Xiaoren Tang, Deborah Levy Marciano, Susan E. Leeman and Salomon Amar. PNAS. April 5, 2005 vol. 102 no. 14, 5132-5137

7. HIV-1-mediated apoptosis of neuronal cells: Proximal molecular mechanisms of HIV-1-induced encephalopathy. Yan Xu, JosephKulkoshy, Roger j. Pomerantz. PNAS. 2004 May 4, 2004 Vol. 101 No. 18.

8. A novel method for high-throughput protein profiling from conditioned media and patient’s sera. Ruo-Pan Huang, RuochunHuang, Yan Fan, and Ying Lin. Ana. Biochem. 2001;294(1):55-62.

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