RayBiotech, Inc.
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1. M Dawood F Zurawska U et al Circulation 2007 115 1398 1407 Somatic Excision Demonstrates that c Jun Induces Cellular Migration and Invasion through Induction of Stem Cell Factor Katiyar S Jiao X Wagner E Lisanti MP Pestell RG Mol Cell Biol 2007 27 4 1356 1369 RayBio Mouse Cytokine Antibody Array G series 6 19 Detection of cytokine protein expression in mouse lung homogenates using suspension bead array McDuffie E Obert L Chupka J Sigler R J Inflamm 2006 3 15 doi 10 1186 1476 9255 3 15 Burn Injury Reveals Altered Phenotype in Mannan Binding Lectin Deficient Mice Moller Kristensen M Hamblin MR Theil S Jensenius JC Takahashi K J Invest Dermatol 2007 127 1524 1531 Overexpression of Heme Oxygenase 1 in Murine Melanoma Increased Proliferation and Viability of Tumor Cells Decreased Survival of Mice Was H Cichon T Smolarczyk R Rudnicka D et al Am J Pathol 2006 169 2181 2198 Vanin 1 licenses inflammatory mediator production by gut epithelial cells and controls colitis by antagonizing peroxisome proliferator activated receptor gamma activity Berruyer C Pouyet L Millet V Martin FM et al J Exp Med 2006 13 2817 2827 A Key Regulatory Role for Histamine in Experimental Autoimmune Encephalomyelitis Disease Exacerbation in Histidine Decarboxylase Deficient mice Musio S Gallo B Scabeni S Lapilla M et al J Immunol 2006 176 17 26 Fusokine Interleuikin 2 Interleukin 18 a Novel Potent Innat2. of CD4 lymphocytes into the brain contributes to neurodegeneration in a mouse model of Parkinson disease Brochard V Combadiere B Prigent A Laouar Y et al J Clin Invest 2009 119 182 192 Transferrin derived synthetic peptide induces highly conserved pro inflammatory responses of macrophages Haddad G Belosevic M Mol Immunol 2009 46 576 586 Human AB Serum and Thrombin Activated Platelet Rich Plasma Are Suitable Alternatives to Fetal Calf Serum for the Expansion of Mesenchymal Stem Cells from Adipose Tissue Kocaoemer A Kern S Kluter H Bieback K Stem Cells 2007 25 1270 1278 Neurofibromin deficient Schwann cells secrete a potent migratory stimulus for Nfl mast cells Yang F C Ingram DA Chen S Hingtgen CM etal J Clin Invest 2003 112 12 1851 1861 Telomere dysfunction induces environmental alterations limiting hematopoietic stem cell function and engraftment Ju Z Jiang H Jaworski M Rathinam C et al Nature Med 2007 13 6 742 747 Transvascular delivery of small interfereing RNA to the central nervous system Kumar P Wu H McBride JL Jung K E et al Nature 2007 448 39 43 Expanded Polyglutamine Huntingtin Protein Suppresses the Secretion and Production of a Chemokine CCLS5 RANTES by Astrocytes Chou S Y Weng J W Lai H L Liao F et al J Neurosci 2008 28 13 3277 3290 Tumor Necrosis Factor alpha Mediates Cardiac Remodeling and Ventricular Dysfunction After Pressure Overload State Sun M Chen
3. recombinant protein and antibody Peptide arrays Phosphorylation arrays ELISA EIA ot ek se Just simply send your samples and we will do the assay for you Technology transfer program Have you developed technologies or reagents interested to the scientific and research community RayBiotech can help you commercialize your technologies reagents and dream RayBio Mouse Cytokine Antibody Array G series 6 17 VI Troubleshooting guide Problem Came Weak signal Inadequate detection Check laser power and PMT parameters Inadequate reagent volumes Check pipetters and or improper dilution ensure correct preparation 2 8 Ensure sufficient incubation Short incubation times Time and change sample incubation step to overnight Too low protein concentration Don t make too low dilution in sample Or concentrate sample Improper storage of kit Store kit at suggested temperature High background Excess of biotinylated antibodies Make sure correct amount of antibodies Excess of streptavidin Make sure correct amount of streptavidin Inadequate detection Check laser power And PMT parameters a Insufficient wash Increase wash time and use more wash buffer Bubbles formed during Avoid bubble formation mncubation during incubation Arrays are not completed Completely cover arrays Covered by reagent with solution RayBio Mouse Cytokine Antibody Array G series 6 18 VII Selected References Using RayBiotech Arrays Infiltration
4. BSA Normally it should only give a background reading Data Extraction Tips e Ignore any comet tails e Define the area for signal capture for all spots as 110 120 micron diameter using the same area for every spot e Use median signal value not the total or the mean e Use local background correction also median value e Exclude obvious outlier data in its calculations Using these guidelines along with using PMT brightness and contrast settings that reduce the background as much as possible we get very good results with interassay and intraassay CV lt 20 even with some imperfections in the antibody spots Threshold of significant difference in expression Any gt 1 5 fold increase or lt 0 65 fold decrease in signal intensity for a single analyte between samples provided that both signals are well above background Mean background 2 standard deviations accuracy 95 RayBio Mouse Cytokine Antibody Array G series 6 13 RayBio Mouse Cytokine Antibody Array 6 97 a gt e Ca ne i fs aa y y a EE a E poss NeG NeG 6Ckine AKI Amphireguin Axl BLC Cardiowophint CD77 rosi Pos poss NEG NEG 6Ckins ALK 1 Amphiregulin Axl BrC Cardiotrophin t_ CD27 9 TNF alpha TRANCE TROY TWEAKR VCAM 1 VEGF VEGF R1 VEGF R3 VEGF D 10 TNF alpha TRANCE TROY TWEAKR VCAM 1 VEGF VEGF R1 VEGF R3_ VEGF D Cont RayBio Mouse Cytokine Antibody Array 6 97 oe a y re ee e
5. RII vewe vwe O ok gt RayBio Mouse Cytokine Antibody Array G series 6 15 RayBiotech Inc the protein array pioneer company strives to research and develop new products to meet demands of the biomedical community RayBio s patent pending technology allows detection of over 180 cytokines chemokines and other proteins in a single experiment Our format is simple sensitive reliable and cost effective Products include Cytokine Arrays Chemokine Arrays ELISA kits Phosphotyrosine kits Recombinant Proteins Antibodies and custom services 1 Antibody arrays Cytokine antibody array Human cytokine antibody arrays Mouse cytokine antibody arrays Rat cytokine antibody arrays Pathway or disease focused antibody arrays Inflammation antibody array Angiogensis antibody array Chemokine antibody array Growth factor antibody array MMP antibody array Atherosclerosis antibody array Quantibody arrays for quantitative measurement of cytokine and other protein concentraton Phosphorylation antibody arrays Biotin label based antibody arrays for high density antibody arrays Antibody analysis tool software ELISA Cell based phosphorylation assay Custom antibody arrays Antibody Recombinant protein Peptide Protein arrays EIA ee ee a e RayBiotech also provides excellent custom service 1 Antibody arrays 2 Protein arrays 3 Peptide synthesis RayBio Mouse Cytokine Antibody Array G series 6 16 Production of
6. RayBio Mouse Cytokine Antibody Array G Series 6 Patent Pending Technology User Manual Revised March 6 2009 RayBio Mouse Cytokine Antibody Array G Series 6 Cat AAM CYT G6 4 RayBio Mouse Cytokine Antibody Array G Series 6 Cat AAM CYT G6 8 RayBio Custom Mouse Cytokine Antibody Array Cat AAM CUST RayBio Mouse Cytokine Antibody Array Service Cat AAM SERV Please read the manual carefully before you start your experiment RayBiotech Inc We Provide You With Excellent Protein Array Systems and Service Tel Toll Free 1 888 494 8555 or 770 729 2992 Fax 1 888 547 0580 Website www raybiotech com Email info raybiotech com RayBiotech Inc RayBio Mouse Cytokine Antibody Array G Series 6 Protocol TABLE OF CONTENTS Pe vilocecc cs c er eee How It Works 0 c cece cece cc eeee eee eeeaeees II Materials Provided c cc cece cece eee e ee ees Additional Materials Required 6 III Overview and General Considerations A Preparation of Samples cee eee ee B Handling Glass Chips 0 eee ee C Incubation 00 ccc cece cece cee eee eee eeeees IV Protocol 0 ccc ccc cece cece cee e neces eeanneeeees A Blocking and Incubation 44 B DC O eerie En EEE EE EE V Interpretation of Results 0c cece eee VI Troubleshooting Guide ccc cece eee VII Re
7. act background Normalize the data from different samples Obtain protein level comparison charts among different samples This analysis tool is very simple and affordable which will not only assist in compiling and organizing your data but also reduces your calculations to a copy and paste step Mouse G Series 1000 Mi IIl MA IM Mouse serum sample 1 Mouse serum sample 2 RayBio Mouse Cytokine Antibody Array G series 6 12 If you do not use our RayBio Analysis Tool you can locate the cytokines by referring to the corresponding RayBio Mouse cytokine Antibody Array map page 13 Please keep in mind that G series 1000 consists of two individual arrays Mouse cytokine antibody array II and Mouse cytokine antibody array IV Mouse Cytokine Antibody Array 6 is on a single array Normalization and comparison For biomarker discovery or for analysis of large number of arrays great attention must be paid to the normalization Our antibody array design includes several controls for normalization and comparison of arrays performing in different membranes and different experiments for more information please read the reference 17 Positive control Positive control is biotinylated protein It can be used to normalize the streptavidin incubation step If the positive signals from different array membranes are similar positive control is a simple and effective way for normalization Negative control Negative control is
8. at room temperature for 10 minutes Or wash using slide chamber Rinse the slide with distilled H20 4 Remove water droplets by centrifuge at 1 000 rpm for 3 minutes and then let slide dry completely in air at least 20 minutes protect from light Make sure the slides are absolutely dry before the scanning procedure 5 Image the signals using laser scanner such Axon GenePix using cy3 channel Note we recommend scanning slides right after experiment You also can store the slide at 20 C in dark for several days If you do not have a laser scanner RayBiotech can provide service for you Just simply send your slide to us and we will take care of it RayBio Mouse Cytokine Antibody Array G series 6 11 V Interpretation of Results The following figure shows RayBio Mouse Cytokine Antibody Array G series 1000 probed with different cell culture supernatant The images were captured using laser scanner The biotin conjugated protein produces positive signals which can be used to identify the orientation and to compare the relative expression levels among the different wells The positive signal can also be used to normalize the signal intensities among array membranes in different experiments The signal intensities obtained from laser scanner can simply be imported into our analysis tool The analysis tool will help you Locate your signal intensities to antibody array map Protein list sorting Average signal intensities Subtr
9. d conditioned media or 300 to 400 ul of 2 5 fold diluted serum or plasma or 10 200 ug of total protein for cell lysates and tissue lysates Dilute the lysate at least 10 fold with I X blocking buffer to make a total volume of 300 to 400 ul Make sure there are no bubbles in the wells RayBio Mouse Cytokine Antibody Array G series 6 9 Note The amount of sample used depends on the abundance of cytokines More sample can be used if signals are too weak If signals are too strong the sample can be diluted further 5 Decant the samples from each well and wash 3 times with 600 ul of 1X Wash Buffer I at room temperature with gentle shaking 2 min per wash Dilute 20X Wash Buffer I with H2O Completely remove wash buffer I in each wash step Note avoid solution flowing into neighboring wells 6 Put the glass chip with frame into a box with 1X Wash Buffer I cover the whole glass slide and frame with Wash Buffer I and wash 2 times at room temperature with gentle shaking for 10 min per wash 7 Decant the 1X Wash Buffer I from each well Put the glass chip with frame into the box with 1X Wash Buffer II cover the whole glass slide and frame with Wash Buffer II and wash 2 times at room temperature with gentle shaking for 5 min per wash Remove all of Wash Buffer II in the well Dilute 20X Wash Buffer II with H O 8 Prepare working solution for biotin conjugated antibodies After brief spinning Add 1450 ul of 1x blocking buffer to t
10. e Cytokine Antibody Microarray slides 1 slide with 4 subarrays 1 or 2 slides e Biotin Conjugated Anti Cytokines 1 tube 4 subarrays 1 or 2 tubes 1 500X Fluorescent Dye conjugated Streptavidin cy3 equivalent 1 tube 4 subarrays 1 or 2 tubes 2X Blocking Buffer 5 ml 10 ml 20X Wash Buffer I 30 ml 20X Wash Buffer II 30 ml 2X Cell Lysis Buffer 10 ml RayBio G series antibody array accessory including slide incubation chamber Gasket Protective cover Snap on sides and adhesive film 30 ml tube e Manual Additional Materials Required Orbital shaker Laser scanner for fluorescence detection Aluminum foil Distilled water Plastic box RayBio Mouse Cytokine Antibody Array G series 6 6 Layout of G series Array 6 HI Overview and General Considerations A Preparation of Samples B Use serum free conditioned media if possible C If serum containing conditioned media is required use uncultured media as a negative control sample since many types of sera contain cytokines D For cell lysates and tissue lysates we recommend using RayBio Cell Lysis Buffer to extract proteins from cell or tissue e g using homogenizer Dilute 2X RayBio Cell Lysis Buffer with H2O we recommend adding proteinase inhibitors to Cell Lysis Buffer before use After extraction spin the sample down and save the supernatant for your experiment Determine protein concentration E We recommend using e 50 100 ul of Condit
11. e and Adaptive Immune Stimulator with Decreased Toxicity Acres B Gantzer M Remy C Futin N Accart N et al Cancer Res 2005 65 20 9536 9546 Clara cels impact the pulmonary innate immune response to LPS Elizur A Adair Kirk TL Kelley DG Griffin GL deMello DE Senior RM Am J Physiol Lung Cell Mol Physiol 2007 293 L383 L392 Synergistic Increases in Intracellular Ca2 and the release of MCP 1 RANTES and IL 6 by Astrocytes Treated with Opiates and HIV 1 Tat El Hage N Gurwell JA Singh IN Knapp PE Nath A Hauser KF Glia 2004 50 91 106 Corticotropin Releasing Hormone Receptor 2 Deficient Mice Have Reduced Intestinal Inflammatory Responses Kokkotou E Torres D Moss AC O Brien M et al J Immunol 2006 177 3355 3361 RayBio Mouse Cytokine Antibody Array G series 6 20 Note RayBio is the trademark of RayBiotech Inc Cytokine protein arrays are RayBiotech patent pending technology This product is intended for research only and is not to be used for clinical diagnosis Our produces may not be resold modified for resale or used to manufacture commercial products without written approval by RayBiotech Inc Under no circumstances shall RayBiotech be liable for any damages arising out of the use of the materials Products are guaranteed for three months from the date of purchase when handled and stored properly In the event of any defect in quality or merchantability RayBiotech s liability to buyer for any cla
12. e ee aaa a a ea aa Deis aa a 2 c27L cD30 CD30L CD36 SR B3 _CTLA 4 CXCL16 Decorin Dkk 1_ E Cadherin _EGF__ Eotaxin e a oen ie cr eo cer rac sama Ko Lopin Leptin o nec mo nee neo meo nec neo neo neo neo neo o sea meo neo meo meo nec nec neo neo nec meo Columns A to AB are on a single membrane We also offer Custom Mouse Cytokine Antibody Arrays You can select the cytokines of interest from the following list and we will produce the customized array at an affordable price For more information please visit our website www raybiotech com Note IL 12 reacts both IL 12p40 and IL 12p70 IL 12p70 only recognizes IL 12p70 Abbreviations IP 10 Interferon inducible protein 10 LAP latency associated peptide TGF B1 LIF leukocyte inhibitory factor MMP Matrix Metalloproteinase Pos positive control Neg negative control All other are used standard abbreviations RayBio Mouse Cytokine Antibody Array G series 6 14 RayBio Mouse Custom Array Antibody List Choose from 146 cytokines and other proteins 6Ckine ALK 1 Amphiregulin CD36 CD40 Ligand Chordin Ea E selectin Fastigand Fe gamma Ril T Fractaikine M carcina Galectin7 acsF _ peame specs MAA a res Lymphotactin MIP 1alpha MIP 1gamma MIP 3 alpha MMPS Neprilysin Osteopontin Osteoporotegerin SDF 1alpha MIP 3 beta Pentraxin 3 TSG OO PRAO RANTES sTNF RI sTNF
13. e profiling of macrophages exposed to Porphyromonas gingivalis its LPS or its FimA Zhou Q Desta T Graves DT Amar S Infect Immunity 2005 73 2 935 43 12 Veto like activity of mesenchymal stem cells functional discrimination between cellular responses to alloantigens and recall antigens Rameshwar P Journal of Immunology 2003 171 7 3426 34 13 The promise of cytokine antibody arrays in drug discovery process Huang RP Yang W Yang Y Flowers L et al Expert Opinion on Drug Discovery 2005 9 601 615 RayBio Mouse Cytokine Antibody Array G series 6 4 Here s how it works Array support Sample Incubation of Sample With arrayed antibody 1 2 hrs Supports Cocktail of Biotin Ab K K KX y K KA Incubation with YYY Biotinylated Ab 1 2 hrs Incubation with h rs Labeled a streptavidin Labeled streptavidin vo as a 7 Detection of signals lat Data analysis and graph RayBio Mouse Cytokine Antibody Array G series 6 5 II Materials Provided Upon receipt all components of the RayBio Mouse Cytokine Antibody Array kit should be stored at 20 C At 20 C the kit will retain complete activity for up to 6 months Once thawed the glass chips Fluorescent dye streptavidin and 2X Blocking Buffer should be kept at 20 C and all other component should be stored at 4 C Use within three months after reagents have been thawed Please use within six months of purchase e RayBio Mous
14. ference List ccc ccc cece cece eee e eens WHO N N N A DB Ua N l 12 18 19 Cytokine Antibody Arrays are RayBiotech patent pending technology RayBio is the trademark of RayBiotech Inc RayBio Mouse Cytokine Antibody Array G series 6 1 I Introduction All cell functions including cell proliferation cell death and differentiation as well as maintenance of health status and development of disease are controlled by many genes and signaling pathways New techniques such as cDNA microarrays have enabled us to analyze the global gene expression However almost all cell functions are executed by proteins which cannot be studied by DNA and RNA alone Experimental analysis clearly shows a disparity between the relative expression levels of mRNA and their corresponding proteins Therefore it is critical to analyze the protein profile Currently two dimensional polyacrylamide SDS page coupled with mass spectrometry is the mainstream approach to analyzing multiple protein expression levels gt However the requirement of sophisticated devices and the lack of quantitative measurements greatly limit its broad application Thus no simple cost effective and rapid method of analysis of multiple protein expression levels has been available to researchers until now Our RayBio Mouse Cytokine Antibody Array is the first commercially available protein array system By using the RayBiotech system scientists can ra
15. he Biotin Conjugated Antibody tube Mix gently Note the diluted biotin conjugated antibodies can be stored at 4 C for 2 3 days 9 Add 350 ul of diluted biotin conjugated antibodies to each corresponding well Incubate at room temperature for 2 hours Note incubation may be done overnight at 4 C 10 Wash as directed in steps 5 and then wash 3 times with 600 ul of 1X Wash Buffer II at room temperature with shaking 2 min per wash Completely remove wash buffer II in each wash step RayBio Mouse Cytokine Antibody Array G series 6 10 11 Add 350 ul of 1 500 fold diluted Fluorescent dye conjugated streptavidin after brief spinning add 1 5 ml of Blocking Buffer to Fluorescent dye conjugated streptavidin tube to each subarray Cover the incubation chamber with Adhesive film Cover the plate with aluminum foil to avoid exposure to light or incubate in dark room 12 Incubate at room temperature for 1 to 2 hours Note incubation may be done overnight at 4 C 13 Wash with Wash Buffer I twice as directed in steps 5 B Fluorescence Detection 1 Decant excess Wash Buffer from wells 2 Disassemble the slide out of the incubation frame and chamber 3 Place the whole slide in a 30 ml centrifuge tube provided add enough Wash Buffer I about 20 ml to cover the whole slide and gently shake at room temperature for 10 minutes Decant Wash Buffer I Repeat Wash Buffer I once Wash with Wash Buffer II about 20 ml with gentle shake
16. he glass chip into incubation chamber and incubation frame as shown below Note if you slide has be assembled you can go to step 3 directly Instructions for incubation chamber assembly G Series and Q series arrays Incubation chamber _ Gasket normally attached to chamber Protective Cover can a a be discarded casas Snap on sides RayBio Mouse Cytokine Antibody Array G series 6 g Carefully place slide at bottom of the chamber as shown The slide will adhere somewhat to the bottom Warning the slide is fragile so do not apply more than gentle force to the apparatus While gently holding chamber and slide place side on chamber as shown beginning with bottom flap first Then press the top of the side into grove on chamber and then apply even gentle pressure from one end to the other Repeat this procedure with the other side 3 Add 500 ul 1 X Blocking Buffer into each well and incubate at room temperature for 30 min to block slides Dilute 2X Blocking Buffer with H O Make sure no bubbles are in the well Note only add reagents to wells printed with antibodies 4 Decant Blocking Buffer from each array well Add 350 ul of each sample to each array well Incubate arrays with sample at room temperature for to 2 hours Dilute sample using 1X Blocking Buffer if necessary Note Incubation may be done overnight at 4 C Note We recommend using 300 to 400 ul of undilute
17. im relating to products shall be limited to replacement or refund of the purchase price HiLytePlus is a trademark of AnaSpec Inc This product is for research use only 2009 RayBiotech Inc RayBio Mouse Cytokine Antibody Array G series 6 21
18. ioned media undiluted or e 50 100 ul of 2 fold to 5 fold diluted sera or plasma or e 10 200 ug of total protein for cell lysates and tissue lysates If you experience high background you may further dilute your sample F Handling glass chips e The microarray slides are sensitive do not touch the surface Hold the slides by the edges only e Handle all buffers and slides with powder free gloves e Avoid breaking glass slide e Handle glass chip in clean environment RayBio Mouse Cytokine Antibody Array G series 6 7 G Incubation e Completely cover array area with sample or buffer during incubation and cover the incubation chamber with adhesive film or plastic sheet protector to avoid drying e Avoid foaming during incubation steps e Perform all incubation and wash steps under gentle rotation e Cover the incubation chamber with adhesive film during incubation particularly when incubation is more than 2 hours or 50 ul of sample or reagent is used e Avoid cross contamination from overflowing solution to neighboring wells e Several incubation steps such as step 3 blocking step 4 sample incubation step 9 biotin Ab incubation or step 12 Fluorescent dye streptavidin incubation may be done overnight at 4 C Please make sure to cover the incubation chamber tightly to prevent evaporation IV Protocol A Blocking and Incubation 1 Take the glass chip out from the box Let air dry for 2 hours 2 Assemble t
19. pidly and accurately identify the expression profiles of multiple cytokines in several hours inexpensively The RayBiotech kit G series is a glass chip format The kit provides a highly sensitive approach to simultaneously detect multiple cytokine expression levels from cell culture supernatant patient s serum tissue lysate and other sources The arrays are manufactured using non contact arrayer The experimental procedure is simple and can be performed in any laboratory The signals from G series arrays are detected using a laser scanner Besides the products listed in this manual RayBiotech also provides RayBio Mouse Cytokine Antibody Array G series 2000 for simultaneous detection of 144 mouse cytokines in single experiment RayBiotech also provides RayBio Human Cytokine Antibody Array G series 4000 which is the only product available in the market that can detect 274 human cytokines in single experiment We also provide quantitative antibody arrays which can RayBio Mouse Cytokine Antibody Array G series 6 2 quantitatively measure multiple protein expression in antibody array technology Pathway specific array systems allow investigators to focus on the specific problem and are becoming an increasingly powerful tool in cDNA microarray system RayBiotech s first protein array system known as RayBio Mouse Cytokine Antibody Array is particularly useful compared with the Mouse cytokine cDNA microarray system Besides the abilit
20. tokine Antibody Array G series 6 3 4 Bone Marrow Stroma Influences Transforming Growth Factor B Production in Breast Cancer Cells to Regulate c myc Activation of the Preprotachykinin I Gene in Breast Cancer Cells Oh HS Moharita A Potian JG Whitehead IP et al Cancer Res 2004 64 6327 6336 5 Recombinant Herpes Simplex Virus Type 1 HSV 1 Codelivering Interleukin 12p35 as a Molecular Adjuvant Enhances the Protective Immune Response against Ocular HSV 1 Challenge Osario Y Ghiasi H J Virology 2005 79 6 3297 3308 6 Dysregulated Inflammatory Response to Candida albicans in a C5 Deficient Mouse Strain Mullick A Elias M Picard S Gros P Infect Immunity 2004 72 10 5868 5849 7 Leukotriene By Strongly Increases Monocyte Chemoattractant Protein 1 in Human Monocytes Huang L Zhao A Wong F Ayala JM Cui J Arterioscler Thromb Vascul Biol 2004 24 1783 1788 8 Giroux M Denis F Human CD1d unrestricted NKT cells release chemokines upon Fas engagement Blood 2005 105 2 703 710 9 Monitoring the response of orthotopic bladder tumors to granulocyte macrophage colony stimulating factor therapy using the prostate specific antigen gene as a reporter Wu Q Esuvaranathan K Mahendran R Clin Cancer Res 2004 10 20 6977 84 10 Neuroglial activation and neuroinflammation in the brain of patients with autism p NA Vargas DL Nascimbene C Krishnan C Zimmerman AW Pardo CA Ann Neurol 2005 57 1 67 81 11 Cytokin
21. y to detect protein expression RayBiotech s system is a more accurate reflection of active cytokine levels because it only detects secreted cytokines and no amplification step is needed Furthermore it is much simpler faster environmentally friendlier and more sensitive Simultaneous detection of multiple cytokines undoubtedly provides a powerful tool to study cytokines Cytokines play an important role in innate immunity apoptosis angiogenesis cell growth and differentiation Cytokines are involved in most disease processes including cancer and cardiac diseases The interaction between cytokines and the cellular immune system 1s a dynamic process The interactions of positive and negative stimuli and positive as well as negative regulatory loops are complex and often involve multiple cytokines Reference List l LPS induces the interaction of a transcription factor LPS induced TNF a factor and STAT6 B with effects on multiple cytokines Tang X Marciano DL Leeman SE Amar S PNAS 2005 102 14 5132 5137 2 HIV 1 mediated apoptosis of neuronal cells Proximal molecular mechanisms of HIV 1 induced encephalopathy Xu Y Kulkoshy J Pomerantz RJ PNAS 2004 101 18 7071 7075 3 Synergistic increases in intracellular Ca 2 and the release of MCP 1 RANTES and IL 6 by astrocytes treated with opiates and HIV 1 Tat El Hage N Gurwell JA Singh IN Knapp PE Nath A Hauser KF GLIA 2005 50 2 91 106 RayBio Mouse Cy
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