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Frequently Asked Questions

  • Q: What is AntibodyArrayTM?

    A: AntibodyArray is a recently developed proteomic tool that allows researchers to exam a large number of proteins simultaneously. In AntibodyArraysTM, a large number of antibodies are arrayed and immobilized on a solid support in a pre-determined order, so that each antibody can be identified later by its position. The immobilized antibodies on the support retain their capabilities of interacting with respective antigens. They recognize and capture the antigens during the incubation with a protein sample. The antigens and their associated protein(s) captured by the arrays can then be studied.

     

  • Q: What problems in my research can AntibodyArrayTM help to solve?

    A: Since AntibodyArraysTM detects proteins, the real players of biological functions, they directly address many difficult questions encountered in biomedical research and drug development. For example, AntibodyArrayTM technology is a powerful but easy-to-use tool for identifying protein-protein interactions, protein posttranslational modifications and protein expressions. The technology can be used to develop an understanding of molecular mechanisms underlying a physiological or pathological process. It can also be used in screening drug candidates.

     

  • Q: What advantages does AntibodyArrayTMtechnology have over other methods, such as yeast two-hybrid screening, in identifying protein-protein interactions?

    A: AntibodyArraysTM are much faster, less expensive and less subjective to false positive results than yeast two-hybrid screening. More importantly, AntibodyArrayTMtechnology can detect protein-protein interactions that only occur in the presence of additional cellular factors or only after posttranslational modifications, which yeast two-hybrid screening is not capable of, because those additional factors or modifications do not exist in yeast.

     

  • Q: How long does it take to perform an AntibodyArrayTM assay?

    A: Unlike some other methods, such as yeast two-hybrid screening that employs complicated procedures and takes months to perform, the AntibodyArrayTM assay takes less than 6 hrs and use a simple procedure similar to Western blotting.

     

  • Q: How sensitive is AntibodyArrayTM technology?

    A: AntibodyArrayTM technology is as sensitive as any other immunological methods. Our customers have routinely detected proteins interacting with low-copy-number cellular proteins.

     

  • Q: What makes AntibodyArrayTM technology sensitive and specific?

    A: The high sensitivity of AntibodyArrayTM technology is due to several reasons. First, we carefully selected the antibodies and used the best ones available. Second, the antibodies are at high concentration on the solid support and the high density significantly increases the binding of antigens. Third, because both capturing and detecting antibodies have to bind the same protein complex in order to produce a signal, the chance for false positive result is much lower compared to that in regular immunoassays that only require one antibody binding.

     

  • Q: Can I reuse an AntibodyArrayTM?

    A: Yes, you can re-probe a used array with a different antibody. Stripping is possible too. Please call Technical Services at Hypromatrix for more information.

     

  • Q: Do I have to purchase expensive equipment to use AntibodyArrayTM?

    A: No. Unlike DNA arrays that require specialized reader to collect and analyze data, AntibodyArrayTM is for routine laboratory use. If you have done a Western blotting, you can perform an AntibodyArrayTM assay.

     

  • Q: How is AntibodyArrayTM different from other "protein chip" technology?

    A: One big difference is that AntibodyArrayTM was developed with laboratory researchers in mind. It requires no expensive special equipment.


Technical Contact

Please call 508-856-7900 or email contact@hypromatrix.com

Publications Citing Hypromatrix AntibodyArrays

Publications Citing Hypromatrix AntibodyArrays


Protein-protein interaction

 

1.       Sakaguchi M, Miyazaki M, Takaishi M, Sakaguchi Y, Makino E, Kataoka N, Yamada H, Namba M, Huh NH. (2003) S100C/A11 is a key mediator of Ca(2+)-induced growth inhibition of human epidermal keratinocytes. J Cell Biol. 163(4):825-35.

 

2.       D Chen, LG Xu, L Chen, L Li, Z Zhai, HB Shu (2003) NIK is a component of the EGF/heregulin receptor signaling complexes. Oncogene, 22, 4348–4355.

 

3.       Konstantin V. Stoletovand Bruce I. Terman  (2004) Bmx is a downstream Rap1 effector in VEGF-induced endothelial cell activation. Biochemical and Biophysical Research Communications 320: 7075.

 

4.       Wu P, Wang P. (2004) Per-Arnt-Sim domain-dependent association of cAMP-phosphodiesterase 8A1 with IkappaB proteins. Proc Natl Acad Sci U S A. 101 (51): 17634-9.

 

5.       Liang M, Liang YY, Wrighton K, Ungermannova D, Wang XP, Brunicardi FC, Liu X, Feng XH, Lin X. (2004) Ubiquitination and proteolysis of cancer-derived Smad4 mutants by SCFSkp2. Mol Cell Biol; 24 (17):7524-37.

 

6.       Betty Y. Zhou  and Johnny J. He (2004) Proliferation inhibition of astrocytes, neurons, and non-glial cells by intracellularly expressed human immunodeficiency virus type 1 (HIV-1) Tat protein. Neuroscience Letters. Volume 359, Issue 3, Pages 155-158

 

7.       Gomez-Santos C, Barrachina M, Gimenez-Xavier P, Dalfo E, Ferrer I, Ambrosio S. (2005) Induction of C/EBP beta and GADD153 expression by dopamine in human neuroblastoma cells. Relationship with alpha-synuclein increase and cell damage. Brain Res Bull; 65(1):87-95.

 

8.       Urtz N, Olivera A, Bofill-Cardona E, Csonga R, Billich A, Mechtcheriakova D, Bornancin F, Woisetschlager M, Rivera J, Baumruker T. Early activation of sphingosine kinase in mast cells and recruitment to FcepsilonRI are mediated by its interaction with Lyn kinase. Mol Cell Biol. 24 (19): 8765-77.

 

9.       Cubelos, B., Giménez, C. and Zafra, F. (2005) The scaffolding protein PSD-95 interacts with the gycine transporter GLYT1 and impairs its internalization. J Neurochem. 95:1047-58.

 

10.   D Stoll, MF Templin, J Bachmann, TO Joos (2005) Protein microarrays: Applications and future challenges. Current Opinion in Drug Discovery and Development 8:239-252.

 

11.   HS Duffy, I Iacobas, DC Spray, AW Ashton (2005) Using Antibody Arrays to Detect Protein-Protein Interactions. Practical methods in Cardiovascular Research. 916-935. Springer Berlin Heidelberg.

 

12.   Chunsik Lee, Johan DixeliusÅsa Thulin, Harukiyo KawamuraLena Claesson-Welsh and Anna-Karin Olsson (2006) Signal transduction in endothelial cells by the angiogenesis inhibitor histidine-rich glycoprotein targets focal adhesions. Experimental Cell Research Volume 312, Issue 13, Pages 2547-2556.

 

13.   Andreas Böttcher, Alfred Böttcher, Gerd Schmitz and Petra Schling (2006) Angiotensin-converting enzyme signalling in human preadipocytes and adipocytes. Central European Journal of Biology Publisher: Central European Science Journals, co-published with Springer-Verlag GmbH ISSN: 1895-104X (Paper) 1644-3635.

 

14.   G M Pierantoni, C Rinaldo,  F Esposito, M Mottolese, S Soddu and A Fusco (2006) High Mobility Group A1 (HMGA1) proteins interact with p53 and inhibit its apoptotic activity Cell Death and Differentiation 13, 1554–1563.

 

15.   Veríssimo F, Silva E, Morris JD, Pepperkok R, Jordan P (2006) Protein kinase WNK3 increases cell survival in a caspase-3-dependent pathway. .Oncogene. 25(30): 4172-82.

 

16.   Jer-Yen Yang, Cong S. Zong Weiya Xia, Yongkun Wei, Mohamed Ali-Seyed, Zheng Li, Kristine Broglio, Donald A. Berry, and Mien-Chie Hung (2006) MDM2 Promotes Cell Motility and Invasiveness by Regulating E-Cadherin Degradation. Mol Cell Biol.  26(19): 7269–7282.

 

17.   Jin Xue, Dan Zhou, Hang Yao, Orit Gavrialov, Melanie J. McConnell, Bruce D. Gelb, and Gabriel G. Haddad. (2007) Novel functional interaction between Na+/H+ exchanger 1 and tyrosine phosphatase SHP-2.Am J Physiol Regul Integr Comp Physiol 292: R2406-R2416, 2007.

 

18.   Wilhelm, Imola; Nagyoszi, Peter; Farkas, Attila E.; Couraud, Pierre-Olivier; Romero, Ignacio A; Weksler, Babette; Fazakas, Csilla; Dung, Ngo Thi Khue; Bottka, Sandor; Bauer, Hannelore; Bauer, Hans-Christian; Krizbai, Istvan A. (2008) Hyperosmotic stress induces Axl activation and cleavage in cerebral endothelial cells. Journal of Neurochemistry. 107(1): 116-126.

 

19.   Josema Torres & Fiona M. Watt  (2008) Nanog maintains pluripotency of mouse embryonic stem cells by inhibiting NFkappaB and cooperating with Stat3. Nature Cell Biology 10, 194 – 201.

 

20.   Yiyu Zou, Wen-Bin Tsai, Chien-Jui Cheng, Chiun Hsu, Young Min Chung, Pao-Chen Li, Sue-Hwa Lin5 and Mickey CT Hu (2008) Forkhead box transcription factor FOXO3a suppresses estrogen-dependent breast cancer cell proliferation and tumorigenesis.  Breast Cancer Research, 10:R21.

 

21.   Degang Wang, Jianyin Long, Fangyan Dai, Min Liang, Xin-Hua Feng and Xia Lin (2008) BCL6 Represses Smad Signaling in Transforming Growth Factor-β Resistance. Cancer Research 68, 783-789.

 

22.   Gonzalez-Gonzalez, Inmaculada M.; Garcia-Tardon, Noemi; Cubelos, Beatriz; Gimenez, Cecilio; Zafra, Francisco (2008) The glutamate transporter GLT1b interacts with the scaffold protein PSD-95. Journal of Neurochemistry. 105(5):1834-1848.

 

 

Protein phosphorylation/Acetylation/modification

 

1.       Abdollahi A, Hahnfeldt P, Maercker C, Grone HJ, Debus J, Ansorge W, Folkman J, Hlatky L, Huber PE. (2004) Endostatin's antiangiogenic signaling network. Mol Cell. 13(5):649-63.

 

2.       Xiaoning Si, Maziar Rahmani, Ji Yuan and Honglin Luo (2005) Detection of Cardiac Signaling in the Injured and Hypertrophied Heart. In Molecular Cardiology: Methods and Protocols. Page 291-303. Edited by Zhongjie Sun.

 

3.       Chunsik Lee, Johan DixeliusÅsa Thulin, Harukiyo KawamuraLena Claesson-Welsh and Anna-Karin Olsson (2006) Signal transduction in endothelial cells by the angiogenesis inhibitor histidine-rich glycoprotein targets focal adhesions. Experimental Cell Research Volume 312, Issue 13, Pages 2547-2556.

 

4.       Koichiro Wada, Atsushi Nakajima, Kazufumi Katayama, Chiho Kudo, Atsuhito Shibuya, Naoto Kubota, Yasuo Terauchi, Masashi Tachibana, Hiroyuki Miyoshi, Yoshinori Kamisaki, Tadanori Mayumi, Takashi Kadowaki and Richard S. Blumberg (2006)  Peroxisome Proliferator-activated Receptor γ-mediated Regulation of Neural Stem Cell Proliferation and Differentiation. The Journal of Biological Chemistry, 281, 12673-12681.

 

5.       Zia A. Khan, Juan M. Melero-Martin, Xiao Wu, Sailaja Paruchuri, Elisa Boscolo, John B. Mulliken, and Joyce Bischoff  (2006) Endothelial progenitor cells from infantile hemangioma and umbilical cord blood display unique cellular responses to endostatin Blood, Vol. 108, No. 3, pp. 915-921.

 

6.       Wada K, Arita M, Nakajima A, Katayama K, Kudo C, Kamisaki Y, Serhan CN. (2006) Leukotriene B4 and lipoxin A4 are regulatory signals for neural stem cell proliferation and differentiation. FASEB J. 20(11):1785-92.

 

7.       Seiichi Okabe, Tetsuzo Tauchi, Akihiro Nakajima, Goro Sashida, Akihiko Gotoh, Hal E. Broxmeyer, Junko H. Ohyashiki, Kazuma Ohyashiki. (2007) Depsipeptide (FK228) Preferentially Induces Apoptosis in BCR/ABL-Expressing Cell Lines and Cells from Patients with Chronic Myelogenous Leukemia in Blast Crisis.  Stem Cells and Development. 16(3): 503-514.

 

8.       Bonnie H. Lemster, Joshua J. Michel, David T. Montag, John J. Paat, Stephanie A. Studenski, Anne B. Newman, and Abbe N. Vallejo (2008) Induction of CD56 and TCR-Independent Activation of T Cells with AgingThe Journal of Immunology, 180, 1979 -1990.

 

9.       Harukiyo Kawamura1, Xiujuan Li, Katsutoshi Goishi2, Laurens A. van Meeteren1, Lars Jakobsson1, Stéphanie Cébe-Suarez3, Akio Shimizu2, Dan Edholm1, Kurt Ballmer-Hofer3, Lena Kjellén4, Michael Klagsbrun2, and Lena Claesson-Welsh1 (2008) Neuropilin-1 in regulation of VEGF-induced activation of p38MAPK and endothelial cell organization. Blood, Vol. 112, No. 9, pp. 3638-3649.

 

10.   Ariel Munitz1, Melissa L. McBride1, Joshua S. Bernstein1, and Marc E. Rothenberg (2008) A dual activation and inhibition role for the paired immunoglobulin-like receptor B in eosinophils. Blood, Vol. 111, No. 12, pp. 5694-5703.

 

11.   Richard Shimonkevitz, PhD, Gregory Thomas, BS, Denetta Sue Slone, MD, Michael Craun, MD, Charles Mains, MD, and David Bar-Or, MD (2008) A Diketopiperazine Fragment of Human Serum Albumin Modulates T-Lymphocyte Cytokine Production Through Rap1. J Trauma; 64:35– 41.

 


Protein expression profiling

 

1.       Gómez-Santos, Cristina; Francisco, Roser; Giménez-Xavier, Pol; Ambrosio, Santiago (2007) Dopamine induces TNF[alpha] and TNF-R1 expression in SH-SY5Y human neuroblastoma cells. NeuroReport: - Volume 18 - Issue 16 - pp 1725-1728.


Manual for Capture AntibbodyArray

  For Manual  please click here.

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