RayBiotech, Inc.
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1. 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 Internal control RayBio antibody arrays also include spiking in protein serving as internal control IC The spiking in proteins do not have cross reactivity with protein in the array It can be used to normalize the entire process Negative control Negative control is 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 e Scan all slides at same PMT 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 back2. chip with frame into the box with 1XWash 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 Remove all of Wash Buffer II in the well Dilute 20X Wash Buffer H with H O 8 Prepare working solution for biotin conjugated antibodies After brief spinning add 300 ul ul of Ix blocking buffer to the Biotin Conjugated Antibody tube Mix gently Note the diluted biotin conjugated antibodies can be stored at 4 C for 2 3 days 9 Add 70 ul of diluted biotin conjugated antibodies to each corresponding well Incubate at room temperature for 2 hours Note incubation may be done at 4 C for overnight 10 Wash as directed in steps 5 and then wash 3 times with 150 ul of IX Wash Buffer II at room temperature with shaking 2 min per wash Completely remove wash buffer II in each wash step 11 Add 70 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 RavBio Human adipokine Antibody Array G series 11 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 to 2 hours Note incubation may be done at 4 C for overnight 13 Wash with Wash Buffer I twice as directed in steps 5 B Fluorescence Detection 1 D
3. is the trademark of RayBiotech Inc RavBio Human adipokine Antibody Array G series 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 T 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 f 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 77 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 RavBio Human Cytokine Antibody Array is the first commercially available protein array system 7 By using the RayBiotech system scientists can rapidly and accurately identify the expression profiles of multiple cytokines in several hours inexpensively The RayBiotech kit G series is a glass slide format The kit provides
4. the relative expression levels among the different wells The internal control IC 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 Link the protein to website for more detailed information on the particular protein Protein list sorting Average signal intensities Subtract background Normalize the data from different samples e 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 Wifi Array VI Array VII Array VI Patient Serum 1 Patient Serum 2 Patient Serum 3 Negative Control If you do not use our RayBio Analysis Tool you can locate the cytokines by referring to corresponding RavBio Human cytokine Antibody Array G series map Normalization and comparison RavBio Human adipokine Antibody Array G series 3 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
5. 74 x RavBio Human adipokine Antibody Array G series 20 Note RavBio 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 claim 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 2008 RayBiotech Inc RavBio Human adipokine Antibody Array G series 21
6. CL21 6ckine CCL28 VIC U IL 12 p70 Marapsin Pancreasin U MCP 1 FLRG IL 13 Rat MCP 2 CD14 FIt 3 Ligand IL 13 Ra2 MCP 3 CD23 Fc epsilon RII Follistatin MCP 4 Fractalkine Procalcitonin Calcitonin Prolactin PSA free PSA total P selectin Ubiquitin 1 VCAM 1 VE Cadherin VEGF VEGF R2 CD40 Ligand CEA L L IL L L L L L RAGE RANK RANTES Resistin S 100b VEGF R3 VEGF C VEGF D 11 13 15 16 17 17B 17C 17R GDF 15 MIC 1 RavBio Human adipokine Antibodv Arrav G series 16 SAA 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 274 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 EIA kits Recombinant Proteins Antibodies and custom services 1 Antibody arrays 2 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 Adipokine antibody arrays Antibody analysis tool softwar
7. Dilute sample using IX Blocking Buffer if necessary We strongly recommend including Internal Control in your assay add 100 ul of Blocking Buffer to IC tube mix well and transfer 1 ul of IC to each well 50 to 100 ul of sample Note when transfer IC use 0 1 ul to 2 ul pipettor Note Incubation may be done at 4 C for overnight Note We recommend using 50 to 100 ul of undiluted conditioned media or 50 to 100 ul of original or 2 5 fold diluted serum or plasma or 10 200 ug of protein for cell lysates and tissue lysates Dilute the lysate at least 10 fold with 1X blocking buffer to make a total volume of 50 to 100 ul Make sure there are no bubbles in the wells RavBio Human adipokine Antibody Array G series 10 Note The amount of sample used depends on the abundance of adipokines More of the 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 150 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 each 7 Decant the IX Wash Buffer I from each well Put the glass
8. F Angiogenin DKK 1 HCC 4 CCL16 Angiopoietin 1 DKK 3 Angiopoietin 2 DKK 4 HGF DPPIV CD26 MMP 7 Siglec 9 sTNF RII IL 28A IFN lambda MMP 8 sTNT RI IL29 IFN lambda 1 MMP 9 TACE TARC HVEM DR6 1 309 AR amphiregulin Dtk IL MPIF 1 ITECK CCL25 TGF alpha Axl E Cadherin ICAM 2 IL MSP a Chain TGF beta 1 B7 1 CD80 EDA A2 EGF IFN gamma IL 5 x ICAM 3 IL 5 R alpha NAP 2 TGF beta 2 6 TGF beta 3 EGFR IGFBP 1 IL 6 sR BCMA TNFRSF17 EG VEGF PK1 ae i IL 7 Nidogen 1 Entactin Thyroglobulin Tie 1 IL 8 NrCAM Tie 2 IL 9 beta IG H3 Endoglin IGFBP 4 NRG1 beta 1 HRG1 beta 1 TIM 1 Betacellulin BTC Endostatin NT 3 IGFBP 5 IL 9 R IGFBP 6 Insulin IP 10 bFGF Eotaxin BLC Eotaxin 2 BMP 4 Eotaxin 3 BMP 5 EpCAM TROP1 Leptin R NT 4 Oncostatin M Osteopontin TIMP 4 TNF alpha LAP TGF b1 Osteoprotegerin TNF beta TPO LEPTIN OB TRAIL R1 b NG Erythropoietin R EPO R IL 1 R4 ST2 LH P Cadherin TRAIL R2 BTC E Selectin IL 1 sRI LIF PDGF R alpha TRAIL R3 CA125 Fas Ligand IL 1 sRIl LIGHT Fas TNFRSF6 IL 10 LIMPII SR B2 PDGF R beta TRAIL R4 Trappin 2 Elafin Carbonic Anhydrase IX CA9 Fer RIIB C IL 10 R alpha Lipocalin 2 NGAL Ferritin IL 10 R beta L Selectin PDGF BB TREM 1 TROY Cardiotrophin 1 CT 1 FGF 4 TAI Lvmphotactin PECAM 1 TSH Cathepsin S FGF 6 IL 12 p40 LYVE 1 Platelet Factor 4 TSLP CCL14a HCC 1 PIGF u PAR C
9. J Clin Endocrinol Metab 2008 93 2255 2262 5 Cytokine secretion by human adipocytes is differentially regulated by adiponectin AICAR and troglitazone Sell H Dietze Schroeder D Eckardt K Ecker J Biochem Biophys Res Comm 2006 343 3 700 706 6 Plasma urine and renal expression of adiponectin in human systemic lupus erythematosus Rovin BH Song H Hebert LA Nadasdy T et al Kidney International 2005 68 1825 1833 7 Adiponectin and functional adiponectin receptor 1 are expressed in airway epithelial cells in Chronic Obstructive Pulmonary Disease Miller M Cho JY Pham A Ramsdell J Broide DH J Immunol 2009 182 684 691 8 Sell H Dietze Schroeder D Kaiser U Eckel J MCP I is a potential player in the negative crosstalk between adipose tissue and skeletal muscle Endocrinol 2005 147 5 2458 2467 9 Autocrine Action of Adiponectin on Human Fat Cells Prevents the Release of Insulin Resistance Inducing Factors Dietze Schroeder D Sell H Uhlig M Koenen M Eckel J Diabetes 2005 54 2003 2011 10 Leptin mediated cytokine release and migration of eosinophils Implications for immunopathophysiology of allergic inflammation Wong CK Cheung PFY Lam CWK Eur J Immunol 2007 37 2337 2348 11 Diet induced adiposity alters the serum profile of inflammation in C57BL 6N mice as measured by antibody array Fenton JI Nufiez NO Yakar S Perkins SN et al Diabetes Obesity Metab http dx doi org 10 1111 1463 1326 2008 009
10. Please use within six months of purchase RavBio Human Adipokine Antibody Microarray slides 4 or 8 subarrays in each glass chip Biotin Conjugated Anti Adipokines 1 or 2 tubes 1 tube 4 subarrays 1 500X Fluorescent Dve conjugated Streptavidin Cy3 equivalent 1 tube 2X Blocking Buffer 5 ml 20X Wash Buffer I 30 ml 20X Wash Buffer II 30 ml Internal control powder tube 2X Cell Lysis Buffer 10 ml RavBio G series antibody array accessory including slide incubation chamber Gasket Protective cover Snap on sides and adhesive film e 30 ml tube Manual Additional Materials Required Orbital shaker Laser scanner for fluorescence detection Aluminum foil Distilled water Plastic box RavBio Human adipokine Antibody Array G series 7 Layout of G series LI mm Antibodv LI Antibodv B Arra A l _ LI ii Bi MI E LI mm Bank gt _ LI Blank bl l LI O LILI LILI LIE Barcode I II jI Barcode JITILQU 4 arrays in one glass chip 8 arrays in one glass chip III Overview and General Considerations A Preparation of Samples Use serum free conditioned media if possible If serum containing conditioned media is required use uncultured media as a negative control sample since many types of sera contain cytokines For cell lysates and tissue lysates we recommend using RayBio Cell Lysis Buffer to extract proteins from cell or tissue e g using homogeniz
11. RavBio Human Adipokine Antibody Array G series 1 Patent Pending Technology User Manual Revised February 18 2009 RavBio Human Adipokine Antibody Array G Series 1 Cat AAH ADI G1 4 RavBio Human Adipokine Antibody Array G Series 1 Cat AAH ADI G1 8 RavBio Human Adipokine Antibody Array Service Cat AAH SER ADI G1 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 RavBio Human Adipokine Antibody Array G Series 1 Protocol TABLE OF CONTENTS I AO GUCHON si c cseidrnvdecesikenncasbievededesaeouties 2 How It WOKS uiscceeecscxeuuesaes ceascdeiemeaeecceoks 6 Il Materials Provided seen nn 7 Additional Materials Required 7 III Overview and General Considerations 8 A Preparation of Samples cceee eee 8 B Handling Glass Chips ee 9 CuUIMGUDAnIONG i oicueereseneteansddatdanecadevecesade 9 IV PROUOCOM hss tea a i la 9 A Blocking and Incubation nn 9 B D tection itik tk ae i 12 V Interpretation of Results 2s sen 12 VI Troubleshooting Guide L cece eee 19 VII Selected References Using RayBiotech Arrays 19 Cytokine Antibody Arrays are RayBiotech patent pending technology RavBio
12. 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 RavBiotech also provides RavBio Human Cytokine Antibody Array G series 1000 for detection of 120 human cytokines in single experiment RavBio Human Cytokine Antibody Array G series 2000 for detection of 174 human cytokines in single experiment and RavBio Human Cytokine Antibody Array G series 4000 for detection of 274 human cytokines in single experiment RavBio Human adipokine Antibody Array G series 2 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 RavBio Human Adipokine Antibody Array is particularly useful compared with the human adipokine cDNA microarray system Besides the ability to detect protein expression RayBiotech s system is a more accurate reflection of active adipokine levels because it only detects secreted adipokines and no amplification step is needed Furthermore it is much simpler faster environmentally friendlier and more sensitive Simultaneo
13. e ELISA Cell based phosphorvlation assav Custom antibodv arravs Antibody Recombinant protein Cytokine protein arrays Quantibody arrays for quantitative measurement of cytokine and other protein concentration 10 Phosphorylation antibody arrays 11 Biotin label based antibody arrays for high density antibody arrays 12 EIA 13 Peptide ONDRA RayBiotech also provides excellent custom service 1 Antibody arrays 2 Protein arrays RavBio Human adipokine Antibody Array G series 17 Peptide synthesis Production of recombinant protein and antibody Peptide arrays Phosphorylation arrays ELISA EIA Assay development OMOIDAN HW 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 RavBio Human adipokine Antibody Array G series 18 VI Troubleshooting guide Problem Weak signal Uneven signal Cause Recommendation Inadequate detection Check laser power and PMT parameters Inadequate reagent volumes Check pipetters and or improper dilution ensure correct preparation l MANT Ensure sufficient incubation Short incubation times Time and change s anple incubation step to overnight Too low protein concentration Don t make too low dilution in sample Or concentrate samp
14. ecant excess Wash Buffer from wells 2 Disassemble the slide out of the incubation frame and chamber 3 Place the whole slide in 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 at room temperature for 10 minutes Or wash using slide chamber Rinse the slide with distilled H O 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 as Axon GenebPix using cy3 or green channel Excitation frequency 532 nm 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 we can provide service for you Just simply send your slide to us and we will take care of it V Interpretation of Results RavBio Human adipokine Antibody Array G series 12 The following figure shows RavBio Human Cytokine Antibody Array G series 2000 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
15. er 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 We recommend using o 50 100 ul of Conditioned media undiluted or o 50 100 ul of 2 fold to 5 fold diluted sera or plasma or o 10 200 ug of total protein for cell lysates and tissue lysates Handling glass chips The microarray slides are sensitive do not touch the surface Grip the slides by the edges only Handle all buffers and slides with powder free gloves Avoid breaking glass slide Handle glass chip in clean environment RavBio Human adipokine Antibody Array G series 8 C Incubation 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 Avoid foaming during incubation steps Perform all incubation and wash steps under gentle rotation 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 Avoid cross contamination from overflowing solution to neighboring wells 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 at 4 C for overnight Please ma
16. ground 2 standard deviations accuracy 95 RavBio Human adipokine Antibody Array G series 14 RayBio Human Adipokine Antibody Arrays 1 for simultaneous detection of 62 human adipokines bi me kk a gis insun iao pike peal een TE Serum MCSF MIF MIP 1beta MSP alpha PARC PDGF AA PDGF AB PDGF BB RANTES F Amyloid A j sale A sili TEEN Di iets kejk ied iad isi xeon er ce ee We also offer Custom Human 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 RavBio Human adipokine Antibody Array G series 15 Human Custom Antibody Array List 285 proteins 4 1BB TNFRSF9 CNTF IL 18 R alpha Cripto 1 GITR MIP 1 alpha MIP 1 beta SCFR c GITR Ligand RP Adiponectin Acrp30 CTACK CCL27 MIP 1 delta SDF 1 alpha GM CSF IL MIP 3 alpha SDF 1 beta Adipsin Factor D CTLA 4 lro IL 2 R alpha MIP 3 beta sgp130 AFP CXCL16 DAN Growth Hormom IL 2 R beta MMP 1 Shh N Siglec 5 Decorin HB EG
17. ke sure to cover the incubation chamber tightly to prevent evaporation IV Protocol A Blocking and Incubation l 2 Take the glass chip out from the box Let air dry for 2 hours Assemble the 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 gt Gasket normally attached to chamber Protective Cover can Eea be discarded ESAN Snap on sides gt RavBio Human adipokine Antibody Array G series 9 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 100 ul 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 well and incubate arrays with sample at room temperature for 1 to 2 hours
18. le Improper storage of kit Store kit at suggested temperature 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 Insufficient wash Bubbles formed dunng incubation Arrays are not completed Covered by reagent Work in clean environment Increase wash time and use more wash buffer Avoid bubble formation during incubation Completely cover arrays with solution VII Selected References Using RayBiotech Arrays 1 Protein Array Reveals Differentially Expressed Proteins in Subcutaneous Adipose Tissue in Obesity Skopkova M Penesova A Sell H Radikova Z et al Obesity 2007 15 2396 2406 RavBio Human adipokine Antibody Array G series 19 2 Adipokines oversecreted by omental adipose tissue in human obesity Maury E Ehala Aleksejev K Guiot Y Detry R Vandenhooft A Brichard SM Am J Physiol Endocrinol Metab 2007 293 E656 E665 3 Atrial natriuretic peptide inhibits the production of adipokines and cytokines linked to inflammation and insulin resistance in human subcutaneous adipose tissue Moro C Klimeakova E Lolmede K Berlan M et al Diabetologia 2007 50 1038 1047 4 Adipokine protein expression pattern in growth hormone deficiency predisposes to the increased fat cell size and the whole body metabolic derangements Ukropec J Penesova A Skopkova M Pura M et al
19. lease of MCP 1 RANTES and IL 6 by astrocytes treated with opiates and HIV 1 Tat GLIA 2005 Apr 15 50 2 91 106 4 Bone Marrow Stroma Influences Transforming Growth Factor f Production in Breast Cancer Cells to Regulate c mvc Activation of the Preprotachvkinin I Gene in Breast Cancer Cells Hvun S Oh Anabella Moharita Pranela Rameshwar CANCER RESEARCH 64 6327 6336 September 1 2004 5 Recombinant Herpes Simplex Virus Tvpe 1 HSV 1 Codelivering Interleukin 12p35 as a Molecular Adjuvant Enhances the Protective Immune Response against Ocular HSV 1 Challenge JOURNAL OF VIROLOGY Mar 2005 Vol 79 No 6 6 Dysregulated Inflammatory Response to Candida albicans in a C5 Deficient Mouse Strain Alaka Mullick Miria Elias Serge Picard Philippe Gros Infection and Immunity Oct 2004 p 5868 5876 DOI 10 1128 IAI 72 10 5868 5876 2004 7 Leukotriene B Strongly Increases Monocyte Chemoattractant Protein 1 in Human Monocytes Li Huang Annie Zhao Frederick Wong Julia M Ayala Jisong Cui Arteriosclerosis Thrombosis and Vascular Biology 2004 24 1783 1788 8 Human CD Id unrestricted NKT cells release chemokines upon Fas engagement Martin Giroux and Francois Denis Van Xu Joseph Kulkoshv Roger j Pomerantz Blood prepublished online September 2 2004 DOI 10 1182 blood 2004 04 1537 9 Monitoring the response of orthotopic bladder tumors to granulocvte macrophage colonv stimulating factor therapy using the prostate specific an
20. linical Cancer Research 2004 Mar 15 10 6 2020 8 16 Antibody array generated profiles of cytokine release from THP 1 leukemic monocytes exposed to different amphotericin B formulations Turtinen LW Prall DN Bremer LA Nauss RE Hartsel SC Antimicrobial Agents Chemotherapy 2004 Feb 48 2 396 403 17 The promise of cytokine antibody arrays in drug discovery process R P Huang W Yang D Yang L Flowers I R Horowitz X Cao and R Huang Expert opinion on drug discovery 2005 9 601 615 RavBio Human adipokine Antibody Array G series 5 Here s how it works Array aX support amend l o Sample lt Incubation of Sample i i YYY With arrayed antibody 1 2 hrs Cocktail of ae ilo Biotin Ab KAKX Y K KX Incubation with YYY Biotinylated Ab 1 2 hrs Labeled TIT A streptavidin KU Incubation with YY Y Labeled streptavidin ee gt Detection of signals Data analysis and graph RavBio Human adipokine Antibody Array G series 6 II Materials Provided Upon receipt all components of the RavBio Human Adipokine 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 internal control 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
21. tigen gene as a reporter Wu Q Esuvaranathan K Mahendran R Clinical Cancer Research 2004 Oct 15 10 20 6977 84 10 Neuroglial activation and neuroinflammation in the brain of patients with autism p NA Diana L Vargas Caterina Nascimbene Chitra Krishnan Andrew W Zimmerman Carlos A Pardo Annals of Neurology 2005 Jan 1 DOI 10 1002 ana 203 15 11 Cytokine profiling of macrophages exposed to Porphyromonas gingivalis its LPS or its FimA Qingde Zhou Tesfahun Desta Dana T Graves and Salomon Amar Infection and Immunity IAI 2005 Feb 73 2 935 43 RavBio Human adipokine Antibody Array G series 4 12 Veto like activity of mesenchymal stem cells functional discrimination between cellular responses to alloantigens and recall antigens Rameshwar P Journal of Immunology 2003 Oct 1 171 7 3426 34 13 Cytokine responses elicited in endothelial cells after treatment with a specific toxin Jaya Pandey BioCompare Product Review May 13 2004 14 Proteomic Characterization of the Interstitial Fluid Perfusing the Breast Tumor Microenvironment A Novel Resource for Biomarker and Therapeutic Target Discovery Julio E Celis Pavel Gromov Teresa Cabezon Jos M A Moreira Noona Ambartsumian Kerstin Sandelin Fritz Rank and Irina Gromova Molecular Cellular Proteomics April 2004 11 3 328 39 15 Increased Expression and Secretion of Interleukin 6 in Patients with Barrett s Esophagus Katerina Dvorakova Harinder Garewal C
22. us detection of multiple adipokines undoubtedly provides a powerful tool to study obesity The area of obesity research is getting hotter ever over the past years One of the key driving factor is that adipose tissue is found no longer to be an inert energy storage organ but is emerging as an active participant in regulating physiological and pathologic processes Many soluble factors have been identified from the adipose tissue and are so called as adipocytokines or adipokines But adipokines are also expressed in a number of other tissues and organs Because all of these factors can act in an autocrine paracrine and endocrine manner in the organisms adipokines are thought to serve as mediators linking obesity inflammation immunity and other obesity related diseases gt Without doubt simultaneous detection of multiple adipokines provides a powerful tool to study adipokines References 1 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 2 HIV 1 mediated apoptosis of neuronal cells Proximal molecular mechanisms of HIV 1 induced encephalopathy Yan Xu Joseph Kulkoshy Roger j Pomerantz PNAS 2004 May 4 2004 Vol 101 No 18 RavBio Human adipokine Antibody Array G series 3 3 Synergistic increases in intracellular Ca 2 and the re
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