How nucelus achnages with Microfabrication of the cells

It will be interesting to look into
1. shape and size variation in nucleus on ns-TiOx, SC TiO2, NSL Patterned subtrates
2. What extacly they recomend (must be corealted with the cytoskelton and ceel fetaures)
3. Dioes effect is same on neurons, fibroblast or differenat?
4. Search for other robust explanations

designing cell based assay using FRAP as microfabrication technique

1. there is lot of weightage to the design facile technique for MFs. NSL, PMCS and mask dsign, NSL, profiometry based scarcthing of ns-TiOx; offer gerat possiblity of MFs for functional substrte for cell based assays. Trying all of these will add new dimesnion to MFs techniques towards cell based assya.
2. Writing a review summerising all these techniques offers good future potenatil.
3. FRAP based photobleaching on solid substartes using adsorption on ns-TiOx, will generate photbleach region, those give wider possiblity of ROS generation and susbsequnet cell detachment regions

A Great Day for motivation to young Indian beginners as scientist

एक और नोबेल पर भारत का नाम
लंदन। भारतीय मूल के अमेरिकी वेंकटरमन रामकृष्णन ने नोबेल पुरस्कार जीत कर दुनिया में भारत का नाम रोशन किया है। उन्हें रसायन विज्ञान के क्षेत्र में वर्ष 2009 का नोबेल पुरस्कार मिलेगा। उन्हें इस पुरस्कार के लिए अमेरिकी वैज्ञानिक थामस ए. स्टेट्ज और इजरायल की वैज्ञानिक अदा ई. योनथ के साथ संयुक्त रूप से चुना गया है। इन तीनों को राइबोसोम की संरचना और इसकी कार्यप्रणाली के संबंध में किए गए महत्वपूर्ण अध्ययन के लिए यह सम्मान दिया जाएगा। रायल स्वीडिश एकेडेमी आफ साइंसेज की ओर से यह घोषणा की गई।

एकेडेमी के मुताबिक तीनों वैज्ञानिकों ने राइबोसोम का त्रिविमीय [3-डी] माडल बनाया, जिससे यह पता चला कि किस तरह विभिन्न एंटीबायोटिक और राइबोसोम आपस में जुड़े रहते हैं। वेंकटरमन ने थामस और योनाथ के साथ मिल कर राइबोसोम के बारे में कई उपयोगी जानकारियां इकट्ठी कीं, जिनकी मदद से कई बीमारियों से बचाव के लिए एंटीबायोटिक बनाने में मदद मिली। इन तीनों ने 'एक्स-रे क्रिस्टलोग्राफी' तकनीक से उन लाखों परमाणुओं की गतिविधियों का अध्ययन किया जो हमारे शरीर की कोशिका में प्रोटीन का निर्माण करने वाले राइबोसोम का निर्माण करते हैं।

रसायन विज्ञान के क्षेत्र में इस अद्भुत योगदान के लिए तीनों वैज्ञानिकों को नोबेल पुरस्कार के रूप में 14 लाख अमेरिकी डालर [करीब सात करोड़ रुपये] मिलेंगे। यह रकम तीनों में बराबर-बराबर बंटेगी। वेंकटरमन ने इस पुरस्कार के लिए अपने साथी वैज्ञानिकों और सहयोगियों का आभार जताया है। अति मुदित वेंकटरमन ने कहा, 'मैं अपने प्रखर सहयोगियों, छात्रों और मेरे लैब में काम करने वाले तमाम शोधकर्ताओं का तहे दिल से ऋणी हूं, क्योंकि विज्ञान सामूहिकता से जुड़ा क्षेत्र है।'

वेंकटरमन से पहले छह और भारतीय या भारतीय मूल के वैज्ञानिक प्रतिष्ठित नोबेल पुरस्कार पा चुके हैं। इनमें रवींद्र नाथ टैगोर [साहित्य], सी.वी. रमन [भौतिकी], हरगोविंद खुराना [मेडिसिन व फिजियोलाजी], सुब्रह्माण्यम चंद्रशेखर [भौतिकी], अम‌र्त्य सेन [अर्थशास्त्र] और वी.एस. नायपाल [साहित्य] के नाम शामिल हैं।

अपने मित्रों के बीच 'वेंकी' नाम से लोकप्रिय वेंकटरमन इन दिनों कैंब्रिज, इंग्लैंड के एमआरसी लेबोरेटरी आफ मोलेक्यूलर बायोलाजी के स्ट्रक्चरल स्टडीज डिवीजन के प्रमुख हैं। वह नोबेल पुरस्कार पाने वाले इस लेबोरेटरी के तेरहवें वैज्ञानिक हैं। 69 साल की स्टेट्ज येल यूनिवर्सिटी में मोलेक्यूलर बायोफिजिक्स एंड बायोकेमिस्ट्री विभाग में प्रोफेसर हैं, जबकि योनाथ इजरायल के वेजमैन इंस्टीट्यूट आफ साइंस में स्ट्रक्चरल बायोलाजी विभाग में प्रोफेसर हैं।

चिदंबरम का लाल रामकृष्णन

वेंकटरमन रामकृष्णन का जन्म 1952 में मंदिरों के शहर चिदंबरम, तमिलनाडु में हुआ। 1971 में उन्होंने बड़ौदा विश्वविद्यालय से भौतिकी में स्नातक किया। उसके बाद आगे की पढ़ाई के लिए वह अमेरिका चले गए। ओहायो यूनिवर्सिटी से पीएच.डी करने के बाद 1976-78 तक उन्होंने कैलिफोर्निया विश्वविद्यालय में पढ़ाया। बाद में उन्होंने अमेरिका की नागरिकता हासिल कर ली।

येल यूनिवर्सिटी से पोस्ट डाक्टरल रिसर्च करने के दौरान उन्होंने एंटीबायोटिक में अहम कारक रायबोसोम की संरचना और कार्य प्रणाली पर गहराई से अध्ययन किया। आजकल वह कैम्ब्रिज के मोलक्यूलर बायोलोजी विभाग के वरिष्ठ वैज्ञानिक हैं और उनके कई शोध पत्र अति महत्वपूर्ण पत्रिकाओं में प्रकाशित हो चुके हैं।

21 सितंबर, 2000 को नेचर पत्रिका में रामकृष्णन के दो शोध पत्र प्रकाशित हुए। इनमें पहली बार उन्होंने 30एस रायबोसोम के 3 आगस्ट्रम संरचना की तीन सह ईकाई पर प्रकाश डाला।

Potentails of Microscopy for nanomedicine.........

1. Study mechanism of Protein-NPs corona formation using FRAP could be a nice techniques.
2. Similarly, metal-prtein interaction during plaue formation cna be best kinetically studied by FRAP technique..........what you say???????????

Designing TiOx thin films with active protein coating

1. Many application in clinical fileds relying heavily on metal, metal oxides and composites, procedures in prosthesis (Bone and Dental) need active support of regenerative material. Could we make thin, ns-TiOx depository films that can be deposited on native tissue for crack tolerance or crack resistance?
2. Amalagm (Hg-Silver)filled carries, coated with ns-TiOx thin film may be more reliable? general longevity (imp issue in dentistry) of Amalgam is 24 years, could above ns-Thin film coating can increase this?
3.

5 things needs to studied by NTs

1. Who is more danger Fe2 or Fe3?
2. What is more important to the cells that determobes bocomptaiblity or bioadversity: Bio-Physico-chemical-mechanical
3. Learning from the homing niche of embryonic and homeopoetic stem cells to design artificial sacffoleds formstem cells...........

aah darling stem cells, wana work with you..................

Patient's own stem cells used to cure cancer
Malathy Iyer, TNN 27 July 2009, 01:30am IST
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MUMBAI: For 15 years, Rakesh Singh (name changed) went about his high-pressure job as a senior engineer in a central government firm with a
transplanted kidney.

Daily, he would pop immuno-suppressant pills to prevent his body from rejecting the donated organ. Then, about 18 months ago, he was struck by an "explosive" form of cancer called multiple myeloma — big boils erupted across and within his body, impairing his ability to sign and speak.

Singh's disease put doctors in the city's Jaslok Hospital in a bind about what line of treatment to follow: reducing the immuno-suppressants to let his body fight the cancer cells could have led to a possible rejection of the kidney.

It was then decided to give him a second transplant — of the bone marrow this time — with his own stem cells. Today, Singh is cancer-free and a medical marvel of sorts.

It has been a month since the 49-year-old underwent a PET (positron emission tomography) scan that categorically showed that he had no more cancer cells in his body. He has been breathing easy since then, but the doctors who treated him over 18 months have got a tad busier: they have a big announcement to make to the world.

Papers are being written, archives are being rechecked and old-timers being consulted before the doctors submit their claim to Nephrology Dialysis Transplant, a European journal.

"Singh is the first kidney transplant patient in the world to undergo a stem cell transplant to beat multiple myeloma (a cancer of the plasma cell) after ablative chemotherapy," say nephrologist Dr Madan Bahadur, hematologist Dr Sameer Shah and oncologist Dr Ganpati Bhat.

KEM Hospital's head of nephrology department, Dr V Hase, says, "The Jaslok patient’s case is of great academic interest. Firstly, it is rare for a kidney transplant patient to develop multiple myeloma. Secondly, no transplant patient in India has undergone a stem cell transplant as a rescue mission against cancer.’’

Dr Hase adds that renal failure is a known complication of multiple myeloma. ‘‘So, in the western world, multiple myeloma patients would undergo stem cell transplant first and a renal transplant later,’’ he says. ‘‘But in the Mumbai case, the opposite has happened.’’

As for the man who underwent the Rs 19-lakh rescue, ‘‘life in the last 18 months has been a struggle for survival’’. He smiles as he recalls the difficulty he had in signing papers.

New Inspiring Ideas and my self Qs on my Topic............

1. Study the effect of micropatterning of fibriblast in presence of MMPs to ascertain how the cells reposnds these molecular scissors in presence of their firm microfabricated roots. Strategy and experiment planning is importnat and need deep discussion with experts in the field as this cn unravel the new insight about the role of MMPs in cancer and VLU..........

Biology and chemistry of matrix metalloproteinases (MMPs)

By Jian Cao, M.D. and Stanley Zucker, M.D. *

マトリックスメタロプロテアーゼ（MMP）とTIMPファミリーの構造、機能、およびMMPがかかわる病気についての解説です（英文）。

1. Introduction
2. Tissue Inhibitors of MMPs (TIMPs)
3. Diseases in which MMPs and TIMPs have been incriminated
   1. Participation of MMPs in various aspects of Cancer
   2. Inflammatory diseases
   3. Cardiovascular disease
   4. Lung disease
   5. Central Nervous System disease
   6. Shock Syndromes
   7. Chronic wounds and inflammation of the skin and oral cavity
4. Conclusion
5. References

1. Introduction

Interstitial collagenase, the first MMP family member identified, was initially discovered in experiments designed to explain collagen remodeling in the metamorphosis of a tadpole into a frog¹. Since collagens (numerous types identified) represent the major structural proteins of all tissues and the chief obstacle to cell migration, it was proposed three decades ago that collagenolytic enzymes play pivotal roles in numerous physiologic (fetal development) and pathologic conditions (cancer and arthritis). More recent scientific interest has expanded to include the role of MMPs in numerous disease states involving the cardiovascular, pulmonary, renal, gastrointestinal, musculoskeletal, visual, and hematopoietic systems.

Matrix metalloproteinase (MMPs) belong to the family of zinc endopeptidases collectively referred to as metzincins. The metzincin superfamily is distinguished by a highly conserved motif containing three histidines that bind to zinc at the catalytic site and a conserved methionine that sits beneath the active site ². The metzincins are subdivided into four multigene families: seralysins, astacins, ADAMs/adamalysins, and MMPs. Although our knowledge of the metzincin biology is rapidly expanding, we still do not fully understand how these enzymes regulate biological functions.

The MMP family is comprised of more than 20 related zinc-dependent enzymes that share common functional domains. These enzymes have both a descriptive name generally based on a preferred substrate and a MMP numbering system based on order of discovery (Figure 1). MMPs were initially characterized by their extensive ability to degrade extracellular matrix proteins including collagens, laminin, fibronectin, vitronectin, aggrecan, enactin, tenascin, elastin, and proteoglycans³. More recently, it has been recognized that MMPs cleave many other types of peptides and proteins and have a myriad of other important functions that may be independent of proteolytic activity ⁴. MMPs have distinct but often overlapping substrate specificities, hence leading to the absence of distinct phenotypes in most genetically-engineered mice with knockdown of specific MMPs; MMP14 is the exception.

Domain Structure of MMPs

Figure 1: History and Domain Structure of MMPs. Compilation of MMPs. Date of discovery of MMP and amino acid sequence listed in parentheses.

The basic structure of MMPs is made up of the following homologous domains: 1) signal peptide which directs MMPs to the secretory or plasma membrane insertion pathway; 2) prodomain that confers latency to the enzymes by occupying the active site zinc, making the catalytic enzyme inaccessible to substrates; 3) zinc containing catalytic domain; 4) hemopexin domain which mediates interactions with substrates and confers specificity of the enzymes; and 5) hinge region which links the catalytic and the hemopexin domain (Figure 1). The smallest MMP in size, MMP7 or matrilysin, lacks the hemopexin domain, yet displays specificity in substrate degradation. Additional structural domains and substrate specificities have led to the division of MMPs into subgroups. The membrane-type MMPs contain an additional 20 amino acid transmembrane domain and a small cytoplasmic domain (MMP14, MMP15, MMP16, and MMP24) or a glycosylphosphatidyl inositol linkage (MMP17 and MMP25), which attaches these proteins to the cell surface. MMP2 and MMP9 (referred to as gelatinases based on their substrate preference) contain fibronectin-like domain repeats which aid in substrate binding.

Two sequence motifs are highly conserved in the protein structure of MMPs. The consensus motif HExGHxxGxxH, found in the catalytic domain of all MMPs, contains 3 histidines that coordinate with the zinc ion (Zn) in the active center. The PRCGxPD motif is located in the C-terminal portion of the prodomain of MMPs; coordination of the cysteine residue (C) of this locus with the zinc atom of the active center confers latency to the proenzyme^3,5.

In vivo activity of MMPs is under tight control at several levels. These enzymes are generally expressed in very low amounts and their transcription is tightly regulated either positively or negatively by cytokines and growth factors such as interleukins ( IL-1, IL-4, IL-6), transforming growth factors (EGF, HGF, TGFß), or tumor necrosis factor alpha (TNFα)^6,7. Some of these regulatory molecules can be proteolytically activated or inactivated by MMPs (feedback effect). Post-transcrpitionally, MMP activity is restricted by the latency conferred by the propeptide located in the N-terminal end of the newly synthesized proenzymes. Activation of MMPs following secretion from cells depends on disruption of the prodomain interaction with the catalytic site, which may occur by conformational changes or proteolytic removal of the prodomain. MMPs that contain furin-like recognition domains in their propeptides (MMP11, MT-MMPs, MMP28) can be activated in the trans Golgi network by members of the subtilisin family of serine proteases. With the exception of MMP2, the mechanism for in vivo activation of secreted MMPs is not well understood. MMP14 plays an integral role in the activation of proMMP2 on the cell surface. Extracellular proteolytic activation of secreted MMPs can be mediated by serine proteases such as plasmin, which implies an interdependence of these two enzyme groups in ECM remodeling ⁷. Some active MMPs can activate other proMMPs e.g. MMP3 activation of MMP9 and MMP1. Once activated, MMPs are further regulated by endogenous inhibitors, autodegradation, and selective endocytosis. Endocytosis of MMP2,9, and 13 through a low density lipoprotein receptor-related protein (LRP) mechanism has been demonstrated⁸. Following secretion from the cell, MMP9 is able to bind to the cell surface where it is somewhat protected from local inhibitors.

2. Tissue inhibitors of MMPs (TIMPs)

The Tissue Inhibitors of MetalloProteinases comprise a four-member family of homologous MMP inhibitors (TIMP1, 2, 3, and 4) ³ (Figure 2). TIMP concentrations generally far exceed the concentration of MMPs in tissue and extracellular fluids, thereby limiting their proteolytic activity to focal pericellular sites. In contrast to the usual inhibitory role, low concentration of TIMP2 enhance MMP14 induced activation of MMP2 by forming a triplex with these proteins on the cell surface ⁷. In addition, TIMPs have been shown to have growth promoting activities which are independent of their MMP inhibitory function and apoptosis-inducing properties (TIMP3). The transcription of TIMPs is regulated by similar cytokines and growth factors that control MMP expression i.e. TGFß, TNFα, IL-1, IL-6, although often in distinctive ways. Other endogenous inhibitors include plasma protein α₂macroglobulin, and a surface inhibitor of MMPs, the RECK inhibitor ⁹.

TIMP Structure

Figure 2: Schematic representation of TIMP-1 structure. TIMP1 contains 12 cysteine residues which form six loop structures through disulfide bonds. The N-terminus of TIMPs 1-4 binds to the catalytic domain of most activated MMPs and inhibits function. The C-terminus of TIMP1 and TIMP2 binds to the hemopexin domain of proMMP2 and proMMP9, respectively; this binding regulates MMP function.

3. Diseases in which MMPs and TIMPs have been incriminated

Thousands of papers have been written on the subject of MMPs dealing with various aspects of their physiologic and pathologic roles in biologic processes. The simplicity of thinking about MMPs solely as extracellular matrix degrading enzymes and TIMPs solely as inhibitors of these processes has been eroded by the recognition of numerous other important roles for these proteins. Our understanding of the biology of MMPs and TIMPs has been expanded with the availability of specific MMP and TIMP knockout mice^,10. Families with genetic mutations of MMP2 have demonstrated important differences between mice and men in terms of mutation phenotypes. The mutated MMP2 phenotype in man looks more like the MMP14 mouse knockout.

3a. Participation of the MMPs in various aspects of Cancer

MMPs are collectively able to degrade virtually all ECM components. In cancer, special emphasis was initially placed on the degradation of type IV collagen, a major protein component of basement membranes by MMP2 and MMP9. Subsequently, it has been demonstrated that many non-ECM proteins can be cleaved by selected MMPs. Deciding which protein substrates are physiologically and pathologically relevant has been difficult to ascertain. MMP-induced release from the cell surface (shedding) of heparin binding epithelial growth factor, insulin-like growth factor, and fibroblast growth factor enhance cell proliferation. On the other hand, release and activation of ECM sequestered TGFß by MMPs can lead to inhibition of cell proliferation. MMP-induced shedding of Fas ligand from the cell surface can either enhance or interfere with cell survival. MMP14 and MMP1 have also been incriminated in enhancing cancer cell migration. MMPs are capable of inducing both positive and negative effects on angiogenesis and immunity. It has been observed that some MMPs readily cleave specific chemokines which may have profound effects on numerous biologic processes, not just limited to inflammation⁴. It has been proposed that cleavage of collagen type IV by MMP2/9 exposes a cryptic site which displays affinity for αvß3 integrin, thereby leading to enhancement of angiogenesis. The capacity of MMPs e.g. MMP12, to cleave plasminogen and generate angiostatin, a powerful inhibitor of tumor angiogenesis in mouse cancer models, reminds us that multi-purpose proteases can display cancer enhancement effects, as well as anti-tumor effects. Clinical experience has taught us that the myriad of MMP functions complicates our ability to predict the outcome of MMP inhibitor use in cancer patients ⁷.

Of considerable interest are the cell types responsible for producing MMPs in cancer. Contrary to expectations, most MMPs in tumors are produced by stromal cells rather than the cancer cells (Figure 3). One explanation for this phenomenon is that cancer cells produce Extracellular Matrix Metalloproteinase Inducer (EMMPRIN), a cell surface glycoprotein, which directly stimulates fibroblasts (through direct cell contact) to produce MMP1, 2, 3, and MMP14 (Figure 2) ¹¹. EMMPRIN is also up regulated in inflammatory cells and has been implicated in tissue injury. The importance of cytokines such as TNF-α, interleukin (IL)-1, and IL-6 in stimulating production of MMPs in disease has been emphasized. Increased levels of TIMP1 and TIMP2 have been identified in malignant stromal tissue.

MMP and Cancer

Figure 3: Schematic diagram of interaction between cancer cells and stromal within a tumor with a focus on role of MMPs.

Based on the important role of MMPs in cancer and successful drug trials in mice, numerous clinical trials were initiated in the 1990’s to test the effectiveness of hydroxamic acid-derived MMP inhibitors (MMPIs) in patients with cancer¹². Based on the lack of effectiveness of several broad spectrum MMPIs in advanced cancer types, these drugs have languished. Retrospective assessment of the design of these clinical trials has led to the recognition that specific MMPIs used in conjunction with cytotoxic chemotherapy in early stage, rather than late stage cancer, needs future consideration.

3b. Inflammatory diseases

Many reports have implicated MMP1,MMP3, and MMP9 in rheumatoid and osteoarthritis. An important role for aggrecanase, a member of the ADAM family of metalloproteinases, in articular damage has been proposed. A clinical trial of an MMPI in arthritis failed to achieve its goal.

3c. Cardiovascular disease

There has been a long-standing interest in the role of MMPs in cardiovascular disease. Numerous studies have demonstrated increased levels of MMPs, especially MMP9, at sites of atherosclerosis and aneurysm formation¹³. The concept that the inflammatory process may play a leading role in the development of atherosclerotic plaques, has led to the suggestion that secretion and activation of MMPs by macrophages induces degradation of extracellular matrix in the atherosclerotic plaque and plaque rupture. Based on these concepts, MMPs have been proposed to represent sensitive markers of inflammation in patients with coronary artery disease. The importance of collecting plasma rather than serum specimens for measuring MMP9 in diagnostic testing has been stressed ¹⁴.

MMP tissue levels are increased in the heart in congestive heart failure. Although inhibitors of MMPs have shown value in experimental models of heart disease, a recent clinical trial demonstrated a lack of efficacy of a broad-spectrum MMP inhibitor in patients with congestive heart failure. The results of this trial emphasize the importance of segregating patients into different pathophysiologic categories that are more likely to respond to a specific drug.

3d. Lung disease

Elevated levels of MMPs have been implicated in the pathophysiology of various lung diseases, including acute respiratory distress syndrome, asthma, bronchiectasis, and cystic fibrosis. MMPs, EMMPRIN, and TIMPs are produced by many of the resident cells in the lung, hence complicating the analysis of their role in disease¹⁵. Potential use of MMP inhibitors for treatment of these disorders in patients remains to be explored.

3e. Central Nervous System disease

Following observations of the critical role of MMP9 in animal models resembling multiple sclerosis and Guillain-Barre’s syndrome, MMPs has been implicated in several different types of neurologic diseases^13,16. Treatment with synthetic inhibitors of MMPs has reversed some of the pathology in animal models of brain injury, especially stroke.

3f. Shock syndromes

MMP8 and MMP9 are stored in the granules of polymorphonuclear leukocytes. These cells are key effectors in inflammatory and infectious processes. A role for these MMPs in shock is supported by studies in MMP9 deficient mice that were shown to be resistant to endotoxic shock. Dubois et al.¹⁷ proposed that specific MMP9 inhibition constitutes a potential approach for the treatment of septic shock syndromes.

3g. Chronic wounds and inflammation of the skin and oral cavity

Acute and chronic wounds are associated with high levels of MMP2 and MMP9. These observations have led to the suggestion that nonhealing ulcers develop an environment containing high levels of activated MMPs, which results in chronic tissue turnover and failure of wound closure¹³. MMP9 has been implicated in blistering skin diseases and contact hypersensitivity^13,18. MMPs have long been implicated in periodontal disease¹⁹ and more recently, in inflammatory bowel diseases. Tetracyclines and chemically-modified tetracyclines, which have inhibitory functions against MMPs, have been shown to be useful in treatment of periodontal disease¹⁹.

4. Conclusion

The involvement of MMPs in disease processes and normal embryologic development continues to be of major interest to scientists from many fields. At present, the pharmaceutical industry remains reluctant to further test MMPIs in human disease, but this might change with the development of more specific inhibitors that lack the troublesome side-effects demonstrated with broad-spectrum MMPIs and a better understanding of the basic role of MMPs in pathophysiology.

5. References

1. Gross J, Lapiere CM. Collagenolytic activity in amphibian tissues; a tissue culture assay. Proc Natl Acad Sci USA. 1962;48:1014-1022.

2. Stoker W, Bode W. Structural features of a superfamily of zinc-endopeptidases: the metzincins. Curr Opin Str Biol. 1995;5:383-390.

3. Nagase H, Woessner F. Matrix metalloproteinases. J Biol Chem. 1999;274:21491-21494.

4. Overall CM, Lopez-Otin C. Strategies for MMP inhibition in cancer: innovations for the post-trial era. Nature Reviews/Cancer. 2002;2:657-672.

5. Birkedal-Hansen H. Proteolytic remodeling of extracellular matrix. Curr Opin Cell Biol. 1995;7:728-735.

6. Sternlicht MD, Werb Z. How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol. 2001;17:463-516.

7. Zucker S, Pei D, Cao J, Lopez-Otin C. Membrane type-matrix metalloproteinases (MT-MMP). in: Cell Surface Proteases. Eds. S. Zucker and W-T Chen. Academic Press. 2003:pp. 1-74.

8. Yang Z, Strickland DK, Bornstein P. Extracellular MMP-2 levels are regulated by the low-density lipoprotein-related scavenger receptor and thrombospondin 2. J Biol Chem. 2001;276:8403-8408.

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Cell based Assays: Future for next generation cell biology

Cell-based assays Support Page

One of the most common uses for the In-Cell Western (ICW) assay is to study the effect of a particular compound or compounds, such as drugs, on a signaling pathway.

The first stage of this is to have an idea of the pathway and the possible effect or effects that a chosen compound may have on this pathway.

In this example, we are looking at the EGFR signaling pathway. Specifically, we will look at 3 targets, p-EGFR, p-STAT3 and p-ERK and the effect of 2 drugs, PD168393 (OR compound 1) and U0126 (OR compound 2) on these targets. It is possible to use the ICW to determine the effect of these drugs on these targets of choice.

Publications

• Quantitative methods for the analysis of protein phosphorylation in drug development
• Epithelial Cell Assays for the Odyssey System
• A High Throughput Drug Discovery for Intracellular Neurodegenerative Disease-Associated Targets using the LI-COR Odyssey Near Infrared System
• Tracking G Protein-coupled Receptor Trafficking Using Odyssey Imaging.