User Manual
Contents
1. Add 100ul standard cytokines or samples to each well Incubate arrays at room temperature for 1 2 hour Longer incubation time is preferable for higher signals Note GCF samples can be either eluted immediately or stored frozen and eluted subsequently with same ratio of sample diluent buffer We recommend using 50 to 100 ul of original or diluted serum plasma conditioned media or other body fluid or 50 500 ug ml of protein for cell and tissue lysates Cover the incubation chamber with adhesive film during incubation if less than 70 ul of sample or reagent is used Note This step may be done overnight at 4 C for best results 8 Wash e Decant the samples from each well and wash 5 times 5 min each with 150 ul of 1x Wash Buffer I at room temperature with gentle Quantibody Human Periodontal Disease Array 1 9 shaking Completely remove wash buffer in each wash step Dilute 20x Wash Buffer I with H O e Optional for Cell and Tissue Lysates Put the glass chip with frame into a box with Ix Wash Buffer I cover the whole glass slide and frame with Wash Buffer I and wash at room temperature with gentle shaking for 20 min e Decant the Ix Wash Buffer I from each well wash 2 times 5 min each with 150 ul of 1x Wash Buffer II at room temperature with gentle shaking Completely remove wash buffer in each wash step Dilute 20x Wash Buffer II with H O D Incubation with detection antibody cocktail and wash 9 Reconstitute the2. QAM CAA 3000 e 120 cytokines QAH CAA 2000 QAM CAA 2000 e 0 cytokines QAH CAA 1000 QAM CAA 1000 e 60 cytokines QAH ANG 1000 QAM CX T Q2000 e 40 cytokines QAH INF 3 QAH CHE 1 QAH GF 1 QAH REC 1 QAH CYT 4 QAH CY T 5 QAH CXT 6 QAH CYT 7 QAM INF 1 QAM CYT 4 QAM CYT 5 QAM CYT 6 e 30 cytokines QAH ANG 2 QAH ANG 3 QAM INT 1000 QAR CYT 3 QAM CHE 1 e 20 cytokines QAH CYT 1 QAH CYT 2 QAM CYT 1 QAM CYT 2 QAM CYT 3 QAM INT 1 QAH TH17 1 QAM TH17 1 e 10 cytokines QAH TH 1 QAH INF 1 QAH INF 2 QAH ANG 1 QAH MMP 1 QAH ADI 1 QAM INT 2 QAR CYT 1 QAR CYT 2 QAR INF 1 QAN CYT 1 QAP CYT 1 QAH IGF 1 e less than 10 cytokines QAH ISO 1 QAH ADI 2 QAP CYT 1 QAM ISO G1 Purpose based selection Custom Arrays e Choose from over 500 cytokine pool Any kind Any number e Order slide only or full service in house e Desired marker not in our pool No problem For certain developmental fee we may be able to add the marker to your panel if the paired antibodies are available on the market Check our website regularly for updated Quantibody products Quantibody Human Periodontal Disease Array 1 20 Note Quantibody is the trademark of RayBiotech Inc Cytokine protein arrays are RayBiotech patent pending technology This product is intended for research only and 1s not to be used for clinical diagnosis Our produces may not be resold modified for resale or used to manufacture commercial products without
3. c 0 00 II General Considerations nn A Preparation of Samples ccc cece eee eens B Handling Glass Chips ee C Incubation nn nn nn nn nem enn IVe PET OUOC Ole a A Complete Air Dry the Glass Chip B Prepare Cytokine Standard Dilutions O o WO N N N ND DW NA W e C Blocking and Incubation 0006 D Incubation with Detection Antibody Cocktaul E Incubation with Cy3 Equivalent Dye Streptavidin 10 sy F Fluorescence Detection eee 11 G Data Analysis 0 cece cece ccc eee cece eee n eee eees 12 V Cytokine Array Map amp Standard Curves 13 VI 8 Point Standards eee nn 14 VII System Recovery 0 ccc cece cece ence eee eee necnes 15 VIII Quantibody Q Analyzer cecceceeceeceueerenees 16 IX Troubleshooting Guide 17 X Select Quantibodv Publications 00000 18 XI Experimental Record Form 0 cece ceca es 19 XII How to Choose Quantibodv Products 20 Quantibody Human Periodontal Disease Array 1 2 I Introduction Periodontal disease 1s a gum disease The symptom ranges from simple gum inflammation gingivitis to periodontitis which results in major damage to the soft tissue and bone that support the teeth In the worst cases teeth are lost Because of the irreversible nature of periodontitis early diagnosis and treatmen
4. detection antibody by adding 1 4 ml of Sample Diluent to the tube Spin briefly 10 Add 80 ul of the detection antibody cocktail to each well Incubate at room temperature for l 2 hour Longer incubation time is preferable for higher signals and backgrounds 11 Decant the samples from each well and wash 5 times with 150 ul of Ix Wash Buffer I and then 2 times with 150 ul of Ix Wash Buffer II at room temperature with gentle shaking Completely remove wash buffer in each wash step E Incubation with Cy3 equivalent dye Streptavidin and wash 12 After briefly spinning down add 1 4 ml of Sample Diluent to Cy3 equivalent dye conjugated streptavidin tube Mix gently 13 Add 80 ul of Cy3 equivalent dye conjugated streptavidin to each well Cover the device with aluminum foil to avoid exposure to light or incubate in dark room Incubate at room temperature for 1 hour Quantibody Human Periodontal Disease Array 1 10 14 Decant the samples from each well and wash 5 times with 150 ul of Ix Wash Buffer I at room temperature with gentle shaking Completely remove wash buffer in each wash step F Fluorescence Detection 15 Disassemble the device by pushing clips outward from the slide side Carefully remove the slide from the gasket Be careful not to touch the surface of the array side 16 Place the slide in the slide Washer Dryer a 4 slide holder centrifuge tube add enough 1x Wash Buffer I about 30 ml
5. l Image scan laser scanner l Data extraction 455 433 443 442 121 122 132 119 2 1 3 2 89 88 90 91 GeneP1x etc ee aeons 55 54 57 56 188 178 189 190 J Data computation Q Analyzer l Final Result pg ml Quantibody Human Periodontal Disease Array 1 12 V Cytokine Array Map amp Standard Curves QAH PDD 1 Standard Curves 1e 5 CRP IFNg IL 1a if W IL 1b 1e 4 A l fy IL 2 omen ae IL 4 IL 6 TA IL 8 A IL 10 1e 3 A Lars 4 IL 12 IL 17 MIP 1a H z MMP 9 1e 2 wy id es MMP 13 OPG Signal Intensity OPN Osteoactivin RANK 1e 1 TGFb1 TNFa 107 10 101 102 103 104 105 106 Concentration pg ml Quantibody Human Periodontal Disease Array 1 13 VI 8 Point Standards After reconstitution of the lyophilized cytokine standard mix the 8 point cytokine concentration used for generating the standard curve of a given antigen is listed below The detection sensitivity of each protein in one experiment is user dependent Try our array specific Quantibody Q Analyzer to see your Limit of Detection LOD Section VIII Serial standard concentration pg ml o s e aera a w o u a e om im 3 o pe ee ea ao pO IA Ce po p e e e a s an IL 4 2 000 oS Ht Wl N See ea a arf ee a m n o u a e om a o we o u a e on o o mes o u a m on m as o E AI TG e p ar f oa ae aer a ox o r
6. slide bag to the back of the slide with a permanent marker before discarding the slide bag Once the slide is disassembled you might not have enough info to distinguish one slide from the other C Incubation Completely cover array area with sample or buffer during incubation 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 lt 70 ul of sample or reagent 1s used Several incubation steps such as step 6 blocking step 7 sample incubation step 10 detection antibody incubation or step 13 Cy3 equivalent dye streptavidin incubation may be done overnight at 4 C Please make sure to cover the incubation chamber tightly to prevent evaporation Quantibody Human Periodontal Disease Array 1 7 IV Protocol A Completely air dry the glass chip 1 Take out the glass chip from the box and let it equilibrate to room temperature inside the sealed plastic bag for 20 30 minutes Remove slide from the plastic bag peel off the cover film and let it air dry at room temperature for another l 2 hours Note Incomplete drying of slides before use may cause the formation of comet tails B Prepare Cytokine Standard Dilutions Note There is only one vial of standard provided in the two slide kit which is enough for making two standard curves Reconstitute the lyo
7. 35 6 829 833 4 Souquiere S et al T Cell tropism of simian T cell leukaemia virus type 1 and cytokine profiles in relation to proviral load and immunological changes during chronic infection of naturally infected mandrills Mandrillus sphinx J Med Primatol 2009 38 4 279 89 5 Sharma et al Induction of multiple pro inflammatory cytokines by respiratory viruses and reversal by standardized Echinacea a potent antiviral herbal extract Antiviral Research 2009 83 2 165 170 6 Altamirano Dimas et al Echinacea and anti inflammatory cytokine responses Results of a gene and protein array analysis Pharmacuetical Biology 2009 47 6 500 508 7 Cheung et al Cordysinocan a polysaccharide isolated from cultured Cordyceps activates immune responses in cultured T lymphocytes and macrophages Signaling cascade and induction of cytokines Journal of Ethonopharmacology 2009 124 1 61 68 8 Du et al P2 380 Identification and characterization of human autoantibodies that may be used for the treatment of prion diseases Alzheimer s and Dementia 2009 4 4 T484 T484 9 Van Rossum et al Granulocytosis and thrombocytosis in renal cell carcinoma a pro inflammatory cytokine response originating in the tumour Neth J Med 2009 67 5 191 4 10 Zhai et al Coordinated Changes in mRNA Turnover Translation and RNA Processing Bodies in Bronchial Epithelial Cells following Inflammatory Stimulation Molecular and Cellular Biolo
8. Quantibodv Human Periodontal Disease Array 1 Quantitative measurement of 20 human periodontal disease associated cytokines Patent Pending Technology User Manual Version Oct 2012 Cat QAH PDD 1 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 ravbiotech com Email info raybiotech com CRP IFNy IL la IL 1B IL 2 IL 4 IL 6 IL 8 IL 10 IL 12 IL 17 MIP la MMP 9 MMP 13 OPG OPN Osteoactivin RANK TGFB1 TNFa One standard glass slide is spotted with 16 wells of identical cytokine antibody arrays Each antibody is arrayed in quadruplicate Cytokine Detected 20 Detection Method Fluorescence with laser scanner Cy3 equivalent dye Sample Volume 50 100 ul per array Reproducibility CV lt 20 See Section V For Array Map e0000000 e0000000 e0000000 e0000000 00000000 00000000 o o o o o o e0000000 e0000000 o0000000 00000000 o0000000 00000000 Fluor dye cy3 equivalent Biotin Streptavidin complex Detect antibody Cytokine Capture antibody Glass Slide Support Quantibody Human Periodontal Disease Array 1 1 TABLE OF CONTENTS lI QVEEeW i TAGE OCUICTION s esaeen roepen en nr Mi ia How It Works srerintenkatirza nien nista saat II Materials Provided ee nnnennn nn mnn Additional Materials Required
9. aaa aor tin 5 row meen OTe Tatas ot 385 Toa ai o TGT 0 BT AZZAR o o aa o me jo IF Quantibodv Human Periodontal Disease Array 1 14 VII System Recovery The antibody pairs used in the kit have been tested to recognize their specific antigen The spiking recovery rate of the cytokines by the kit in 2x diluted Human serum and 2x diluted Human cell culture media CM is listed in the following table The kasi recoverv rate for culture media and serum BU ed e Kos Tara lm mar Tajna mar er a sar aera ar or fm a w ejja mr KM IL 1f 1 000 631 63 513 51 IL 2 2 000 2 536 105 49 2 152 105 IL 4 1 000 1 244 123 10 1 361 135 IL 6 1 000 2 436 7196 2 1 023 100 IL 8 1 000 1 156 100 956 95 IL 10 1 000 726 72 l 590 59 IL 12 1 000 1 198 120 2 857 85 IL 17 5 000 3 461 69 2 3 255 65 MIP 1a 5 000 7 134 140 3 934 79 MMP 9 5 000 2 587 51 3 368 4 067 14 MMP 13 10 000 129 9 919 98 lt 6 128 61 OPN 50 000 47 043 92 203 28 861 57 Osteoactivin 5 000 4 993 98 2 493 4 538 41 RANK 25 000 68 19 119 76 21 17 503 70 Quantibody Human Periodontal Disease Array 1 15 VIII Quantibody Q Analyzer Quantibody Q Analyzer is an array specific Excel based program However it is not a simple calculation macro as it contains sophisticated data analysis Key features e Simplicity Easy to operate and requires no professional training With a simple copy and paste process the cytokine concentration 1s det
10. can then be visualized through the addition of the streptavidin labeled Cy3 equivalent dye using a laser scanner Unlike the traditional ELISA Quantibody products use array format By arraying multiple cytokine specific capture antibodies onto a glass support multiplex detection of cytokines in one experiment is made possible In detail one standard glass slide is spotted with 16 wells of identical cytokine antibody arrays Each antibody together with the positive controls is arrayed in quadruplicate The slide comes with a 16 well removable gasket which allows for the process of 16 samples in one slide Four slide chips can be nested into a tray which matches a standard microplate and allows for automated robotic high throughput process of 64 arrays simultaneously For cytokine quantification the array specific cytokine standards whose concentration has been predetermined are provided to generate a standard curve for each cytokine In a real experiment standard cytokines and samples will be assayed in each array simultaneously through a sandwich ELISA procedure By comparing signals from unknown samples to the standard curve the cytokine concentration in the samples will be determined Quantibody array kits have been confirmed to have similar detection sensitivity as traditional ELISA Our current high density Quantibody kits allow scientists to quantitatively determine the concentration of 320 human or 160 mouse cytokines in a single exp
11. during incubation neighboring wells usage Inadequate standard reconstitution or Reconstitute the lyophilized standard well at Improper dilution the room temperature before making serial Poor standard dilutions Check pipettes and ensure proper curve serial dilutions Inadequate detection Increase laser power that the highest standard concentration for each cytokine receives the highest possible reading yet remains unsaturated Use freeze thawed cytokine standards Always use new cytokine standard vial for new set of experiment Discard any leftover Overexposure Lower the laser power Dark spots Completely remove wash buffer in each Hich wash step 18 Insufficient wash Increase wash time and use more wash background buffer Slide is allowed to dry out Don t dry out slides during experiment Quantibody Human Periodontal Disease Array 1 17 X Select Quantibody Publications 1 Stechova et al Influence of Maternal Hyperglycaemia on Cord Blood Mononuclear Cells in Response to Diabetes associated Autoantigens Scandinavian Journal of Immunology 2009 70 2 149 158 2 Willingham SB et al NLRP3 NALP3 Cryopyrin facilitates in vivo caspase 1 activation necrosis and HMGBI release via inflammasome dependent and independent pathways J Immunol 2009 183 3 2008 15 3 El Karim et al Neuropeptides Regulate Expression of Angiogenic Growth Factors in Human Dental Pulp Fibroblasts Journal of Endodontics 2009
12. eriment This is not only one of the most efficient products on the market for cytokine quantification but makes it more affordable for quantification of large number of proteins Simultaneous detection of multiple cytokines undoubtedly provides a powerful tool for drug and biomarker discovery Quantibody Human Periodontal Disease Array 1 4 How It Works Array support YYYYY li a Samples l o Incubation of Sample yyy With arrayed antibody 1 2 hr Supports Cocktail of 4 Biotin Ab XX M k KK Incubation with 1 2 hr Biotinylated Ab Labeled IN AW AN 4 treptavidi eee K K Incubation with 1 hr vv Cy3 equivalent dye Labeled streptavidin Jose 7 Detection of signals lat Data analysis and graph Quantibody Human Periodontal Disease Array 1 5 II Materials Provided Upon receipt all components of the Quantibody 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 chip cytokine standard mix detection antibody cocktail and Cy3 equivalent dye conjugated Streptavidin should be kept at 20 C and all other components may be stored at 4 C The entire kit should be used within 6 months of purchase Components Item Description 1 Slide kit 2 Slide kit Quantibody Array Glass Chip Sample Diluent 20X Wash Buffer I 20X Wash Buffer II Lyophilized cytokine standard mix Detection antibody cocktail C
13. ermined e Outlier Marking amp Removing The software can automatically mark and remove the outlier spots for more accurate data analysis e Normalization The program allows for intra and inter slide normalization for large number of samples e Iwo Positive Controls The program takes the two positive controls in each array for normalization e Two Analytical Algorithms Users can choose either linear regression or log log algorithms to meet their analytical needs e Two Data Outputs standard curves and digital concentration e User Intervention The program allows for user manual handling of those outliers and other analytical data e Lower and Upper Limits Determination The program automatically marks out the values below or above the detection range e Standard Deviation The program outputs the standard deviations of the quadruplicate spots for data accuracy e Analytical Tips Q Analyzer analysis tips are included in the program Quantibody Human Periodontal Disease Array 1 16 IX Troubleshooting guide Inadequate detection Increase laser power and PMT parameters Inadequate reagent volumes or Check pipettes and ensure correct improper dilution preparation Weak Signal change sample incubation step to overnight sample sample Improper storage of kit Store kit as suggested temperature Don t freeze thaw the slide Arrays are not completed covered by Completely cover arrays with solution Uneven signal reagent film
14. gy 2008 28 24 7414 7426 11 Gao et al A Chinese herbal decoction Danggui Buxue Tang activates extracellular signal regulated kinase in cultured T lymphocytes FEBS Letters 2007 581 26 5087 5093 This reference validates mulitplex ELISA results for several analytes with standard ELISA test results 12 Piganelli et al Autoreactive T cell responses new technology in pursuit of an old nemesis Editorial Review Pediatric Diabetes 2007 8 249 251 Quantibody Human Periodontal Disease Array 1 18 XI Experiment Record Form Date File Name Laser Power PMT Well No Sample Name 2 s Q H ili i E E elelee Quantibody Human Periodontal Disease Array 1 19 XII How to Choose Quantibody Products Species based selection e Human QAH e Mouse QAM e Rat QAR CYT 1 QAR CYT 2 QAR CYT 3 QAR INF 1 e Porcine QAP CYT 1 e Non Human Primates NHP QAN CYT 1 Function based selection e THI TH2 THI17 Array QAH TH 1 QAH TH17 QAM TH17 e Inflammation Arrays QAH INF 1 QAH INF 2 QAH INF 3 QAM INF 1 QAR INF 1 e Angiogenesis Arrays QAH ANG I QAH ANG 2 QAH ANG 3 QAH ANG 1000 e Chemokine Arrays QAH CHE 1 QAM CHE 1 e MMP Array QAH MMP 1 e Immunoglobin Isotype Array QAH ISO 1 QAM ISO G1 Cytokine Number based selection e 320 cytokines QAH CAA 7000 e 280 cytokines QAH CAA 6000 e 240 cytokines QAH CAA 5000 e 200 cytokines QAH CAA 4000 e 160 cytokines QAH CAA 3000
15. notransferase Because of the limited availability of sample volumes most of the previous research only worked on one or several targets The traditional method for cytokine detection and quantification is through the use of an enzyme linked immunosorbent array ELISA While the traditional method works well for a single protein the overall procedure 1s time consuming and requires a lot of sample Take the advantage of advancement in microarray technology over the last decade Raybiotech has pioneered the development of cytokine antibody arrays which has now been widely applied in the research community with hundreds of peer reviewed publications such as in Cell and Nature Quantibody Human Periodontal Disease Array 1 3 Quantibody array our quantitative array platform uses the multiplexed sandwich ELISA based technology and enables researchers to accurately determine the concentration of multiple cytokines simultaneously It combines the advantages of the high detection sensitivity specificity of ELISA and the high throughput of the arrays Like a traditional sandwich based ELISA it uses a pair of cytokine specific antibodies for detection A capture antibody is first bound to the glass surface After incubation with the sample the target cytokine is trapped on the solid surface A second biotin labeled detection antibody is then added which can recognize a different isotope of the target cytokine The cytokine antibody biotin complex
16. philized standard within one hour of usage If you must use the standard for two different days store only the Std1 dilution at 80 C Prepare serial dilution of cytokine standards 100ul 100ul 100ul 100ul 100ul 100ul AS OSOS TS OS GD amp Add 500ul pa Sample Diluent 200ul 200ul 200ul 200ul 200ul 200ul 100ul Vial Labels Stdi Std2 Std3 Std4 Std5 Std6 Std7 CNTRL 2 Reconstitute the Cytokine Standard Mix lyophilized by adding 500u1 Sample Diluent to the tube For best recovery always quick spin vial prior to opening Dissolve the powder thoroughly by a gentle mix Labeled the tube as Std Quantibody Human Periodontal Disease Array 1 8 3 Label 6 clean microcentrifuge tubes as Std2 to Std7 Add 200ul Sample Diluent to each of the tubes 4 Pipette 100ul Stdl into tube Std2 and mix gently Perform 5 more serial dilutions by adding 100ul Std2 to tube Std3 and so on 5 Add 100ul Sample Diluent to another tube labeled as CNTRL Do not add standard cytokines or samples to the CNTRL tube which will be used as negative control For best results include a set of standards in each slide Note Since the starting concentration of each cytokine is different the serial concentrations from Stdl to Std7 for each cytokine are varied which can be found in section VI C Blocking and Incubation 6 Add 100ul Sample Diluent into each well and incubate at room temperature for 30 min to block slides 7 Decant buffer from each well
17. t is critical Clinical measurements include probing pocket depth bleeding on probing clinical attachment loss plaque index and radiographs etc While such methods are useful for the staging of periodontal disease they are only indicators of previous disease status rather than the present disease activity There is a need for the development of new diagnostic tests that can reflect the active disease state which will be useful for disease diagnosis prognosis and monitoring the effectiveness of periodontal therapy Gingival crevicular fluid GCF is a tiny amount bodily fluid transuded from periodontal tissues into the gingival crevice and periodontal pocket The constituents of GCF originate from serum gingival tissues and from both bacterial and host response cells which reflect the biology and physiology of the local tissues Meanwhile GCF could be easily collected by noninvasive means such as paper strips absorbent points and micropipettes As a result proteins in GCF have been the ideal and hot targets pursued for candidate disease specific biomarker research for the last several decades Most analyzed periodontal disease related proteins in GCF are inflammatory cytokines eg IL Ib IL 6 IL 8 IL 10 IL 12 IFNg TNFa and CRP matrix metalloproteinases eg MMP 8 MMP 9 and MMP 13 and their inhibitors TIMPs bone metabolism related cytokines eg OPG OPN RANK and RANKL and enzymes eg alkaline phosphatase and aspartate ami
18. to cover the whole slide and then gently shake at room temperature for 15 minutes Decant Wash Buffer I Wash with Ix Wash Buffer II about 30 ml with gentle and gently shake at room temperature for 5 minutes 17 Remove water droplets completely by one of the following ways e Put the glass chip into the Slide Washer Dryer and dry the glass chip by centrifuge at 1 000 rpm for 3 minutes without cap e Or dry the glass chip by a compressed N stream e Or gently apply suction with a pipette to remove water droplets Do not touch the array only the sides 18 Imaging The signals can be visualized through use of a laser scanner equipped with a Cy3 wavelength such as Axon GenePix Make sure that the signal from the well containing the highest standard concentration Stdl receives the highest possible reading yet remains unsaturated Note In case the signal intensity for different cytokine varies greatly in the same array we recommend using multiple scans with a higher PMT for low signal cytokines and a low PMT for high signal cytokines Quantibody Human Periodontal Disease Array 1 11 G Data Analysis 19 Data extraction can be done with most of the microarray analysis software GenePix ScanArray Express Array Vision or MicroVigene For quantitative data analysis our Quantibody Q Analyzer software is available It gives visual output as well as digital values More information can be found in section VIII Experiments
19. 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 This product is for research use only 2012 RayBiotech Inc Quantibody Human Periodontal Disease Array 1 21
20. y3 equivalent dye conjugated Streptavidin Slide Washer Dryer Adhesive device sealer Manual aay sy 2 3 4 5 6 7 8 9 p lt Co See Section VI for detailed cytokine concentrations after reconstitution Additional Materials Required Orbital shaker Laser scanner for fluorescence detection Aluminum foil Distilled water 1 5ml Polypropylene microcentrifuge tubes Quantibody Human Periodontal Disease Array 1 6 HI General Considerations A Preparation of Samples GCF collection was most commonly performed using paper strips inserted into the gingival crevice until mild tissue resistance is felt A sampling period of 30s has been adopted by most investigators Subsequently the volume of GCF collected on the paper strip 1s quantified by an electronic device eg Periotron 8000 GCF samples can be either eluted immediately or stored frozen and eluted subsequently with sample diluent buffer We recommend the following parameters for other samples 50 to 100 ul of original or diluted serum plasma cell culture media or other body fluid or 50 500 ug ml of protein for cell and tissue lysates B Handling glass chips Do not touch the surface of the slides as the microarray slides are very sensitive Hold the slides by the edges only Handle all buffers and slides with latex free gloves Handle glass chip in clean environment Because there 1s no barcode on the slide transcribe the slide serial number from the
Download Pdf Manuals
Related Search