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1. VI Troubleshooting guide Weak signal or no 1 Taking too much time 1 The whole Detection process must be signal for Detection completed in 30 min 2 Film developer does 2 Fix film developer not work properly 3 Did not mix HRP 3 Mix tube containing 1 000X HRP streptavidin well before Conjugated Streptavidin well before use since use precipitation may form during storage 4 Sample is too dilute 4 Increase sample volume e g using undilute sample or using more cells e g seed 2 million cells After 1 or 2 days change complete medium with low serum medium and collect conditioned medium 2 day later Use about 1 to 2 ml of conditioned medium for experiment 5 Other 1 Reduce blocking concentration by diluting in 1 X Wash Buffer II 3 Slightly increase biotin antibody concentrations 4 Expose film for overnight to detect weak signal Uneven signal 1 Bubbles formed 1 Remove bubble during incubation during incubation 2 Membranes were not 2 Completely cover membranes with solution completely covered by solution High background 1 Exposure to x ray file 1 Decrease exposure time is too long 2 Membranes were 2 Completely cover membranes with solution 3 Sample is too 3 Use more diluted sample concentrated RayBio Human Cytokine Antibody Array Protocol 17 VII Selected References Using RayBiotech Antibody Arrays 1 Protein Array Reveals Differentially Expressed Proteins in Subcu2. MCP 1 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 RayBio Human Cytokine Antibody Array Protocol 18 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 Secreted proteins and genes in fetal and neonatal pig adipose tissue and stromal vascular cells Hausman GJ Poulos SP Richardson RL Barb CR et al J Animal Science 2006 84 1666 1681 12 Diet induced adiposity alters the serum profile of inflammation in C57BL 6N mice as measured by antibody array Fenton JI Nu ez NO Yakar S Perkins SN et al Diabetes Obesity Metab http dx doi org 10 1111 1463 1326 2008 00974 x RayBio Human Cytokine Antibody Array Protocol 19 RayBio Human Cytokine Antibody Array Protocol 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 RayBio
3. cell functions including cell proliferation cell death and differentiation as well as maintenance of health status and development of disease are controlled by a multitude of genes and signaling pathways New techniques such as cDNA microarrays have enabled us to analyze global gene expression However almost all cell functions are executed by proteins which cannot be studied simply through DNA and RNA techniques Experimental analysis clearly shows a disparity between the relative expression levels of mRNA and their corresponding proteins Therefore analysis of the protein profile is critical Currently two dimensional polyacrylamide SDS page coupled with mass spectrometry 15 the mainstream approach to analyzing multiple protein expression levels However the requirement of sophisticated devices and the lack of quantitative measurements greatly limit their broad application Thus effective study of multiple protein expression levels has been complicated costly are time consuming until now Our RayBio Human Cytokine Antibody Array is the first commercially available cytokine antibody array system Applying similar principle new we have developed Human Cytokine Antibody Arrays By using the RayBiotech system scientists can rapidly and accurately identify the expression profiles of multiple adipokines in several hours inexpensively The RayBiotech kit provides a simple array format and highly sensitive approach to simulta
4. fold diluted HRP conjugated streptavidin e g add 2 ul of HRP conjugated streptavidin to 1998 ul 1X Blocking Buffer to each membrane Note mix the tube containing 1 000X HRP Conjugated Streptavidin well before use since precipitation may form during storage 10 Incubate at room temperature for 2 hours Note incubation may be done at 4 C for overnight 11 Wash as directed in steps 4 and 5 RayBio Human Cytokine Antibody Array Protocol 10 B Detection Do not let the membrane dry out during detection The detection process must be completed within 40 minutes without stopping 1 Proceed with the detection reaction Add 250 ul of 1X Detection Buffer C and 250 ul of 1X Detection Buffer D for one membrane mix both solutions Drain off excess wash buffer by holding the membrane vertically with forceps Place membrane protein side up mark is on the protein side top left corner on a clean plastic sheet provided in the kit Pipette the mixed Detection Buffer on to the membrane and incubated at room temperature for 2 minute Ensure that the detection mixture is completely and evenly covers the membrane without any air bubbles 2 Drain off any excess detection reagent by holding the membrane vertically with forceps and touching the edge against a tissue Gently place the membrane protein side up on a piece of plastic sheet mark is on the protein side top left corner Cover another piece of plastic sheet on
5. to the array protocol particularly during your first experiment Typical results using Obesity Antibody arrays Human Adipokine Antibody Arrays 1 x e e e e e Patient s Serum Control By comparing the signal intensities relative expression levels of adipokines can be made The intensities of signals can be quantified by densitometry Positive control can be used to normalize the results from different membranes being compared The signals also can be detected and quantitated by using a chemiluminescence imaging device The RayBio Analysis Tool is a program specifically designed for analysis of RayBio Obesity Antibody Arrays This tool will not only assist in compiling and organizing your data but also reduces your calculations to RayBio Human Cytokine Antibody Array Protocol 12 a copy and paste Call RayBiotech Inc at 770 729 2992 for ordering information If you do not use our RayBio Analysis Tool you can locate the cytokines by referring to corresponding RayBio Human cytokine Antibody Array G series map 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 16 in introd
6. PTIN OB PARC TRAIL R1 Erythropoietin R EPO R BTC E Selectin CA125 Fas Ligand IL 1 sRIl LIGHT Fas TNFRSF6 Li P Cadherin TRAIL R2 H LIF PDGF R alpha TRAIL R3 PDGF R beta TRAIL R4 LIMPII SR B2 Trappin 2 Elafin Fcr RIIB C IL 10 R alpha Lipocalin 2 NGAL TREM 1 Carbonic Anhydrase IX CA9 Ferritin IL 10 R beta L Selectin PDGF BB TROY Cardiotrophin 1 CT 1 FGF 4 IL 11 Lymphotactin PECAM 1 TSH Cathepsin S FGF 6 IL 12 p40 LYVE 1 CCL14a HCC 1 FGF 7 IL 12 p70 Marapsin Pancreasin CCL21 6ckine FGF 9 FLRG IL 13 IL 13 Rat Platelet Factor 4 TSLP PIGF u PAR Procalcitonin Calcitonin Ubiquitin 1 Prolactin VCAM 1 Fit 3 Ligand IL 13 Ra2 PSA free VE Cadherin CD23 Fc epsilon RII Follistatin IL 15 ku PSA total VEGF CD27 Fractalkine IL 16 P selectin VEGF R2 Galectin 7 IL 17 IL 17B IL 17F RAGE VEGF R3 MDC RANK VEGF C RANTES VEGF D Resistin IL 17R IL 17C MICA S 100b RayBio Human Cytokine Antibody Array Protocol 14 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 15 simple sensitive reliable and cost effective Products include Cytokine Arrays Chemokine Arrays ELISA kits Phosphotyrosine kits EIA kits Recombinant Protein
7. RayBio Human Obesity Antibody Array 1 User Manual Revised February 4 2009 RayBio Human Obesity Antibody Array 1 Cat AAH ADI 1 RayBio Custom Human Cytokine Antibody Array AAH CUST RayBio Human Cytokine Antibody Array Service Cat AAH SERV Please read manual carefully before starting experiment We Provide You with Excellent Protein Array System 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 Human Obesity Antibody Array Protocol TABLE OF CONTENTS IntrodHclObu sessi RELIEF a ndun 2 6 II Materials 1 7 Additional Materials Required 7 III Overview and General Considerations 8 A Preparation of 8 B Handling Array 8 C iT ANON PP 9 IV M L KK KK 9 A Blocking and Incubation 9 D 10 V Interpretation of Re sultS L EEE 11 Troubleshooting I7 VII Reference List 18 Cytokine protein arrays are RayBiotech patent pending technology RayBio is the trademark of RayBiotech Inc RayBio Human Cytokine Antibody Array Protocol 1 I Introduction All
8. fied 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 Without doubt simultaneous detection of multiple adipokines provides a powerful tool to study adipokines 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 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 GLIA 2005 Apr 15 50 2 91 106 RayBio Human Cytokine Antibody Array Protocol 3 4 Bone Marrow Stroma Influences Transforming Growth Factor p Production in Breast Cancer Cells to Regulate c myc Activation of the Preprotachykinin I Gene in Breast Cancer Cells Hyun S Oh Anabella Moharita Pranela Rameshwar CANCER RESEARCH 64 6327 6336 September 1 2004 5 Recombinant Herpes S
9. implex Virus Type 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 CD1d unrestricted cells release chemokines upon Fas engagement Martin Giroux and Francois Denis Yan Xu Joseph Kulkoshy 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 granulocyte macrophage colony stimulating factor therapy using the prostate specific antigen 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 20315 Cytokine responses elicited in endothelial cells afte
10. in 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 RayBio Human Cytokine Antibody Array Protocol 13 www raybiotech com Human Custom Antibody Array List 285 pr 4 1BB TNFRSF9 CNTF es IL 18 R alpha ACE 2 Cripto 1 CRP Activin A oteins GITR GITR Ligand MIP 1 alpha SCF IL 18 R beta MIP 1 beta IL 1ra MIP 1 delta SDF 1 alpha Adiponectin Acrp30 CTACK CCL27 Adipsin Factor D CTLA 4 AFP CXCL16 GM CSF IL 2 R alpha MIP 3 beta sgp130 IL 2 R beta MMP 1 Shh N IL 2 MIP 3 alpha SDF 1 beta AgRP ART DAN IL 2 gamma MMP 2 Siglec 5 ALCAM Decorin HB EGF IL 21 R MMP 3 Siglec 9 Angiogenin DKK 1 Angiopoietin 1 HCC 4 CCL16 hCGa intact IL 22 MMP 7 sTNF RII IL 28A IFN lambda sTNT RI Angiopoietin 2 Bate2 M EGF IFN gamma IL 6 NCAM 1 BCAM EGFR IGFBP 1 IL 6 sR BCMA TNFRSF17 EG VEGF PK1 BDNF ENA 78 beta IG H3 Endoglin ICAM 3 HGF M 1 2 IGFBP 3 IL29 IFN lambda 1 TACE IL 5 R alpha NAP 2 TGF beta 2 TGF beta 3 NGF R Thyroglobulin IL 7 Nidogen 1 Entactin Tie 1 IL 8 NrCAM Tie 2 NRG1 beta 1 HRG1 beta 1 Betacellulin BTC Endostatin IGFBP 4 IGFBP 5 NT 3 4 BMP 4 Eotaxin 3 IGF I sR Oncostatin M TIMP 4 11 Osteopontin TNF alpha BMP 5 EpCAM TROP1 LAP TGF b1 Osteoprotegerin TNF beta ErbB3 LE
11. in complete activity for up to 6 months Once thawed the array membranes and 1X Blocking Buffer should be kept at 20 C and all other component should be stored at 4 C After thawing the reagents the kit must be used within three months and please use the kit within six months of purchase e RayBio Human Obesity Antibody Array membranes 2 4 8 membranes e Biotin Conjugated Anti Adipokines 1 2 4 tubes each tube for two membranes 1 000X HRP Conjugated Streptavidin 50 ul 1 X Blocking Buffer 25 50 ml 20X Wash Buffer I 10 20 ml 20X Wash Buffer II 10 20 ml 2X Cell Lysis Buffer 10 20 ml Detection Buffer C 1 5 2 5 ml Detection Buffer D 1 5 2 5 ml Eight Well Tray 1 each Manual Additional Materials Required Small plastic boxes or containers Orbital shaker Plastic sheet protector or SaranWrap Kodak X Omat AR film REF 165 1454 and film processor or Chemiluminescence imaging system RayBio Human Cytokine Antibody Array Protocol 7 III Overview and General Considerations A Preparation of Samples Use serum free conditioned media if possible If serum containing media is required use an uncultured media aliquot 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 homogenizer Dilute 2X RayBio Cell Lysis Buffer with H O we recommend adding prote
12. inase 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 per membrane o 1l ml of Conditioned media undiluted or o 1l ml of 2 fold to 5 fold diluted sera or plasma or o 50 500 ug of total protein for cell lysates and tissue lysates use 200 250 ug of total protein for first experiment Dilute the lysate at least 10 fold with 1 X blocking buffer Note The amount of sample used depends on the abundance of adipokines More of the sample can be used if the signals are too weak If the signals are too strong the sample can be diluted further If you experience high background you may further dilute your sample B Handling Array Membranes Always use forceps to handle membranes and grip the membranes by the edges only Never allow the array membranes to dry during experiments RayBio Human Cytokine Antibody Array Protocol 8 C Incubation e Completely cover the membranes with sample or buffer during incubation and cover the eight well tray with lid to avoid drying e Avoid foaming during incubation steps e Perform all incubation and wash steps under gentle rotation e Several incubation steps such as step 2 blocking step 3 sample incubation step 8 biotin Ab incubation and step 11 HRP streptavidin incubation may be done at 4 C for overnight but make sure to cover the 8 well plate tightly
13. neously detect multiple adipokine expression levels from conditioned media patient s sera cell lysate tissue lysates and other sources Traditionally adipokines are detected by using ELISA However RayBiotech s approach has several advantages over ELISA Most importantly our approach can detect many adipokines simultaneously with trace amount of sample RayBio Human Cytokine Antibody Array Protocol 2 Pathway specific array systems allow investigators to focus on the specific problem and are becoming an increasingly powerful tool in cDNA microarray systems RayBiotech s first antibody array system known as Human obesity antibody array 15 particularly useful in comparison 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 Simultaneous 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 15 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 identi
14. r treatment with a specific toxin Jaya Pandey BioCompare Product Review May 13 2004 12 Mlinar B et al Molecular mechanisms of insulin resistance and associated diseases Clinica Chimica Acta 2007 375 20 35 RayBio Human Cytokine Antibody Array Protocol 4 13 Tilg H et al Adipocytokines mediators linking adipose tissue inflammation and immunity Nat Rev Immunol 2006 6 772 783 14 Trujillo ME et al Adipose tissue derived factors impact on health and disease Endocr Rev 2006 27 762 78 15 Van Gaal LF et al Mechanisms linking obesity with cardiovascular disease Nature 2006 444 875 880 e 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 RayBio Human Cytokine Antibody Array Protocol 5 Here s how it works Array support j YYY a a Samples M YY with arrayed antibody YYY supports 1 2 hrs Cocktail of X K i Biotin Ab KK Incubation with Y Y Biotinylated Ab 1 2 hrs streptavidin X jq Incubation with vY Y labeled Streptavidin 1 hrs ce Detection of signals Data analysis and graph RayBio Human Cytokine Antibody Array Protocol 6 II Materials Provided Upon receipt all components of the RayBio Human Obesity Antibody Array kit should be stored at 20 C to 80 C At 20 C to 80 C the kit will reta
15. s 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 Obesity antibody arrays Antibody analysis tool software ELISA Cell based phosphorylation assay Custom antibody arrays 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 RayBiotech also provides excellent custom service 1 Antibody arrays RayBio Human Cytokine Antibody Array Protocol 15 Protein arrays Peptide synthesis Production of recombinant protein and antibody Peptide arrays Phosphorylation arrays ELISA EIA Assay development 00 1 O0 tA N 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 Human Cytokine Antibody Array Protocol 16
16. taneous Adipose Tissue in Obesity Skopkova M Penesova A Sell H Radikova Z et al Obesity 2007 15 2396 2406 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 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
17. tech 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 Kodak X Omat is the trademark of Eastman Kodak Company This product is for research use only 2009 RayBiotech Inc RayBio Human Cytokine Antibody Array Protocol 21
18. the array Gently smooth out any air bubble Avoid using pressure on the membrane 3 Expose the array to x ray film we recommend to use Kodak X Omat AR film and detect the signal using film developer or the signal can be detected directly from the membrane using a chemiluminescence imaging system Expose the membranes for 40 Seconds Then re expose the film according to the intensity of signals If the signals are too strong background too high reduce exposure time e g 5 30 seconds If the signals are too weak increase exposure time e g 5 20 min or overnight Or re incubate membranes overnight with 1x HRP conjugated streptavidin and redo detection in the second day 4 Save membranes in 20 C to 80 C for future reference RayBio Human Cytokine Antibody Array Protocol 11 V Interpretation of Results The following figure shows RayBio Human Obesity Antibody Array membranes probed with conditioned media from two different cell lines Membranes were exposed to Kodak X Omat film at room temperature for minute The biotin conjugated IgG produces positive signals which can be used to identify the orientation and to compare the relative expression levels among the different membranes One important parameter is background To obtain the best results we suggest that several exposures be attempted We also strongly recommend using a negative control which the sample is replaced with an appropriate mock buffer according
19. to prevent evaporation IV Protocol A Blocking and Incubation 1 Place each membrane into the provided eight well tray means the antibody printed side 2 Add 2 ml 1X Blocking Buffer and incubate at room temperature for 30 min to block membranes Make sure there are no bubbles between the membranes 3 Decant Blocking Buffer from each container and incubate membranes with 1ml of sample at room temperature for 1 to 2 hours Dilute sample using 1X Blocking Buffer if necessary Note Incubation may be done at 4 C for overnight 4 Decant the samples from each container and wash 3 times with 2 ml of 1X Wash Buffer I at room temperature with shaking Please allow 5 min per wash Dilute 20X Wash Buffer I with H O 5 Wash 2 times with 2 ml of 1X Wash Buffer II at room temperature with shaking Allow 5 min per wash Dilute 20X Wash Buffer II with H5O RayBio Human Cytokine Antibody Array Protocol 9 6 Prepare working solution for primary antibody Add 100 ul of Ix blocking buffer to the Biotin Conjugated Anti Adipokines tube Mix gently and transfer all mixture to a tube containing 2 ml of 1 blocking buffer Note the diluted biotin conjugated antibodies can be stored at 4 C for 2 3 days 7 Add 1 ml of diluted biotin conjugated antibodies to each membrane Incubate at room temperature for 1 2 hours Note incubation may be done at 4 C for overnight 8 Wash as directed in steps 4 and 5 9 Add 2 ml of 1 000
20. uction Positive control Positive control is biotinylated protein It can be used to normalize the streptavidin incubation step If the positive signals from different arrays are similar positive control is a simple and effective way for normalization Negative control Negative control is BSA Normally it should only give a background reading Human Adipokine Antibody Arrays 1 for simultanous detection of 62 human adipokine s MCSF MIF MIP 1beta MSP alpha i _SDF 1 sTNT RI j SDF 1 sTNF RII TECK STNT RI TGF beta TIMP 1 TIMP 2 TNF alpha TGF beta TIMP 1 TIMP 2 TNF alpha PDGF AA VEGF VEGF PDGF AB XEDAR XEDAR PDGF BB RANTES BLANK BLANK BLANK BLANK Resistin BLANK BLANK 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Hormone d Fas crow HCC 4 gamma IGFBP 1 terae erap a IGF I IGF I SR IL 1 R4 ST2 IL 1 sRI IL 10 IL 11 Hormone e IL 12 IL 1alpha IL 1beta IL 6 IL 6 sR 12 10 LeptinR Leptin LIF Lymphotactin 1 MCP 3 f IL 12 IL 1alpha IL 1beta IL 6 IL 6 sR 12 10 LeptinR Leptin LIF Lymphotactin 1 MCP 3 g MCSF MIF MIP ibeta MSP alpha PDGF BB RANTES Resistin Serum Amyloid A Serum Amyloid A POS POS Abbreviations OSM oncostatin M TPO thrombopoietin Pos positive control Neg negative control All others use standard abbreviations RayBiotech also offers customized cytokine prote
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