Performance characteristics, continued

Expected values Allopregnanolone Competitive ELISA Kit High Low A number of -extracted serum samples from pregnant Expected Conc. Observed Conc. % Catalog Number EIAALLO (96 tests) Sample Sample humans were tested in the assay. Adjusted neat of (pg/mL) (pg/mL) % % Recovery Pub. No. MAN0018776 Rev B.0

allopregnanolone was >2,900 pg/mL. A number of serum samples from non-pregnant human samples were extracted and tested in 80 20 3,441.7 3,493.3 101.5 Note: For safety and biohazard guidelines, see the “Safety” appendix in the ELISA Technical Guide (Pub. no. MAN0006706). Read the Safety the assay. Adjusted neat concentrations of allopregnanolone 60 40 2,691.7 2,811.0 104.4 Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. ranged from 738 to >1,698 pg/mL. Urine samples, diluted 1:4 to 40 60 1,941.7 1,860.0 95.8 1:16 from pregnant and non-pregnant human and other Product description 20 80 1,191.6 1,078.3 90.5 mammalian species were tested in the assay. Adjusted The Allopregnanolone ELISA Kit is a solid-phase monoclonal antibody based competitive Enzyme-Linked Immunosorbent Assay (ELISA). of allopregnanolone varied from 1,500 to >46,000 Mean Recovery 98.1% This assay is designed to detect and quantify the level of allopregnanolone in extracted serum, plasma, and dried fecal samples, or urine pg/mL for non-pregnant and 7,700 to >57,000 pg/mL for pregnant and tissue culture media. The assay recognizes allopregnanolone independent of species. samples. Specificity The following cross reactants were tested in the assay and Sensitivity calculated at the 50% binding point. Contents and storage The analytical sensitivity of allopregnanolone is 129.7 pg/mL. This Kit and components are shipped at –20°C. Upon receipt, store the kit at –20°C. Once open, store the kit at 4°C and use within 2 weeks. Cross-reactivity % was determined by adding two standard deviations to the mean Store the Allopregnanolone Conjugate at –20°C or lower after opening. O.D. obtained when the zero and Std7 were assayed 19 times, and Allopregnanolone 100 calculating the corresponding concentration. 2.19 Components Quantity

Linearity Tetrahydrodeoxycorticosterone (THDOC) 3.08 Allopregnanolone Standard; 100,000 pg/mL allopregnanolone 125 µL Linearity was determined for fecal extracts and diluted urine by Dihydrodeoxycorticosterone (DHDOC) <0.08 Assay Buffer Concentrate (5X) 28 mL taking two samples, one with a low level and one with a higher 0.095 level of allopregnanolone, and mixing them in the ratios given Antibody Coated Wells, 96-well strip-well plate coated with goat anti-mouse IgG 1 plate <0.08 below. The measured concentrations were compared to the Allopregnanolone Antibody 3 mL expected values based on the ratios used. 5α-dihydroprogesterone <0.08 Fecal Extract Corticosterone <0.08 Allopregnanolone Conjugate 3 mL High Low <0.08 Expected Conc. Observed Conc. % Wash Buffer Concentrate (20X) 30 mL Sample Sample (pg/mL) (pg/mL) Recovery 0.12 % % TMB (Tetramethylbenzidine) Substrate 11 mL 11α-hydroxyprogesterone <0.08 80 20 2,046.9 2,027.5 99.1 20α-hydroxyprogesterone <0.08 Stop Solution; contains 1 M HCl, CAUSTIC 5 mL 60 40 1,625.1 1,580.4 97.2 <0.08 Plate Sealer 1 40 60 1,203.4 1,187.6 98.7 <0.08 20 80 781.7 764.3 97.8 Estradiol <0.08 Materials required but not supplied Prepare 1X Wash Buffer Mean Recovery 98.2% 1. Dilute 15 mL of Wash Solution Concentrate (20X) with <0.08 • Distilled or deionized water 285 mL of deionized or distilled water. Label as 1X Wash • Microtiter plate reader with software capable of measurement at or near 450 nm (preferably with correction between 570 nm Buffer.

and 590 nm). 2. Store the concentrate and 1X Wash Buffer in the refrigerator. Limited product warranty Use the diluted buffer within 3 months. • Plate washer–automated or manual (squirt bottle, manifold Life Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies' General Terms and dispenser, or equivalent) Conditions of Sale found on Life Technologies' website at www.thermofisher.com/us/en/home/global/terms -and-conditions.html . If Prepare 1X Assay Buffer • Calibrated adjustable precision pipettes and glass or plastic you have any questions, please contact Life Technologies at www.thermofisher.com/support. 1. Dilute 14 mL of Assay Buffer (5X) with 56 mL of deionized or tubes for diluting solution distilled water. Label as 1X Assay Buffer.

Consult 2. Store the concentrate and 1X Assay Buffer in the refrigerator. Catalog Batch Temperature Caution, consult Procedural guidelines Use by Manufacturer instructions for accompanying documents 1X Assay Buffer is stable at 4°C for 3 months. Number code limitation use IMPORTANT! Reagents are lot-specific. Do not mix or Manufacturer: Life Technologies Corporation | 7335 Executive Way | Frederick, MD 21704 | USA interchange different reagent lots from various kit lots. The information in this guide is subject to change without notice. • Review the Procedural guidelines and Plate washing

DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, directions in the ELISA Technical Guide available at MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT. thermofisher.com. Important Licensing Information: This product may be covered by one or more Limited Use Label Licenses. By use of this product, you accept the terms and conditions of all applicable Limited Use Label Licenses. • Allow reagents to reach room temperature before use. Mix to © 2020 Thermo Fisher Scientific Inc. All rights reserved. MAGPIX, FLEXMAP 3D, xPONENT, Luminex, Luminex 100, and Luminex 200 are trademarks of Luminex Corporation. Excel redissolve any precipitated salts. is a trademark of Microsoft Corporation. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. • Solutions containing sodium azide will inhibit the activity of the peroxidase conjugate. Ensure that there is no contamination of labware or the plate washer with azide containing solutions.

thermofisher.com/support | thermofisher.com/askaquestion For research use only. Not for use in diagnostic procedures. thermofisher.com 01 October 2020 Sample preparation guidelines Perform ELISA (Total assay time: 2.5 hours) • Refer to the ELISA Technical Guide at thermofisher.com for detailed sample preparation procedures. IMPORTANT! Perform a standard curve with each assay. • Collect samples in pyrogen/endotoxin-free tubes. • Freeze samples after collection if samples will not be tested immediately. Avoid multiple freeze-thaw cycles of frozen samples. Thaw Allow all components to reach room temperature before use. Mix all liquid reagents prior to use. completely and mix well (do not vortex) prior to analysis. Determine the number of 8-well strips required for the assay. Insert the strips in the frames for use. Re-bag any unused strips and frames, • Avoid the use of hemolyzed or lipemic sera. and store desiccated at 2°C to 8°C for future use. The silica pack in the bag keeps the plate dry, and turns from blue to pink if the bag is not properly sealed. • If large amounts of particulate matter are present in the sample, centrifuge or filter sample prior to analysis. Bind antigen Prepare samples a. Add 50 μL of standards or samples (see “Prepare samples” on page 2) to the appropriate wells. Sample concentrations should be within the range of the standard curve. Because conditions may vary, each investigator should determine b. Add 75 μL of 1X Assay Buffer into wells for detecting non-specific binding (NSB). the optimal dilution for each application. c. Add 50 μL of 1X Assay Buffer into maximum binding wells (B0 or Zero standard). Use all samples within 2 hours of dilution, or store at –20°C or lower until ready to perform assay. d. Add 25 μL of Allopregnanolone Conjugate to each well. Sample type Procedure e. Add 25 μL of Allopregnanolone Antibody to each well except NSB wells. f. Tap the side of the plate to mix. Cover the plate with plate sealer. Serum and plasma 1. Add diethyl ether or ethyl acetate to samples at a 5:1 (v/v) solvent:sample ratio. g. Shake at room temperature for 2 hours. If the plate is not shaken signals bound will be ~ 45% lower. 2. Mix solutions by vortexing for 2 minutes. Allow solvent layer to separate for 5 minutes. OR 3. Freeze samples in a dry ice/ethanol bath and pour solvent solution from the top of the sample into a Shake the plate in a plate shaker at room temperature for 15 minutes to ensure adequate mixing of the clean tube. Repeat steps 1-3 for maximum extraction efficiency, combining top layer of ether solutions. reagents. Incubate at 4°C for 16-18 hours. 4. Dry pooled solvent samples down in a Speedvac for 2-3 hrs, or under a stream until dry. If h. Thoroughly aspirate the solution and wash wells 4 times with 300 μL of 1X Wash Buffer. samples need to be stored they should be kept desiccated at -20°C. Add chromogen 5. Redissolve samples at room temperature in 1X Assay Buffer. Use a minimum of 125 μL of Assay Buffer. a. Add 100 μL TMB Substrate to each well. The substrate solution will begin to turn blue. Urine Dilute samples ≥1:4 with 1X Assay Buffer. b. Incubate for 30 minutes at room temperature without shaking. Note: A Urinary Creatinine Detection Kit (Cat. no. EIACUN) is available for measuring urine creatinine for Note: TMB should not touch aluminum foil or other metals. normalization of allopregnanolone in a random urine specimen. Add stop solution Dried feces See detailed extraction protocol on the product page at thermofisher.com Add 50 μL Stop Solution to each well. Tap side of the plate gently to mix. The solution in the wells changes from Note: The ethanol concentration in the final diluted Assay Buffer dilution added to the well should be <5%. blue to yellow. Tissue culture media Dilute sample with 1X Assay Buffer. The investigator should determine the appropriate dilution for media samples prior to running the assay.

Dilute standards Note: Use glass or plastic tubes for diluting standards. Read the plate and generate the standard curve The Allopregnanolone Standard contains an organic solvent. Pipette the standard up and down several times to wet the pipet tip before 1. Read the absorbance at 450 nm. Read the plate within 10 minutes after adding the Stop Solution. transfer to ensure that volumes are accurate. 2. Average the duplicate Optical Density (OD) values for each standard and sample. Use curve-fitting software to generate the standard curve. A four parameter algorithm provides the best standard curve fit. Optimally, the background absorbance may be subtracted 1. Add 50 µL Allopregnanolone Standard to one tube containing 450 µL 1X Assay Buffer and label as 10,000 pg/mL allopregnanolone. from all data points, including standards, unknowns and controls, prior to plotting. 2. Add 250 µL Standard Diluent Buffer to each of 7 tubes labeled as follows: 5000, 2500, 1250, 625, 312.5, 156.25, and 0 pg/mL 3. Calculate the concentrations for unknown samples and controls from the %B/B0 curve. Multiply value(s) obtained for sample(s) by allopregnanolone. the appropriate factor to correct for the sample dilution. 3. Make serial dilutions of the standard as described below in the dilution diagram. Mix thoroughly between steps. Note: Dilute samples producing signals lower than that of the highest standard in 1X Assay Buffer and reanalyze. Multiply the 4. Use the standards within 2 hours of preparation. concentration by the appropriate dilution factor.

Performance characteristics Intra-assay precision Standard curve (example) Samples were assayed in replicates of 20 to determine precision within an assay. The following data were obtained for the various standards over the range of 0–10,000 pg/mL allopregnanolone. Parameters Sample 1 Sample 2 Sample 3 Note: 1 pg/mL of allopregnanolone is equivalent to 3.14 pM. Mean (pg/mL) 3,576.5 1,768.8 545.7 Std Allopregnanolone (pg/mL) Net Optical Density (450 nm) %B/B0 %CV 3.7 4.6 12.9 10,000 0.118 12.37 CV = Coefficient of Variation 5,000 0.229 24.00 Inter-assay precision 2,500 0.396 41.51 Samples were assayed in duplicates in 20 assay runs by four 1,250 0.583 61.11 operators to determine precision between assays. 625 0.721 75.58 Parameters Sample 1 Sample 2 Sample 3 312.5 0.849 88.99 Mean (pg/mL) 3,538.8 1,710.6 499.2 156.25 0.885 92.77 %CV 6.2 7.0 11.2 0 0.954 100 CV = Coefficient of Variation

Note: The NSB gave a mean OD value of 0.063.

2 ELISA Kit Product Information Sheet ELISA Kit Product Information Sheet 3 Sample preparation guidelines Perform ELISA (Total assay time: 2.5 hours) • Refer to the ELISA Technical Guide at thermofisher.com for detailed sample preparation procedures. IMPORTANT! Perform a standard curve with each assay. • Collect samples in pyrogen/endotoxin-free tubes. • Freeze samples after collection if samples will not be tested immediately. Avoid multiple freeze-thaw cycles of frozen samples. Thaw Allow all components to reach room temperature before use. Mix all liquid reagents prior to use. completely and mix well (do not vortex) prior to analysis. Determine the number of 8-well strips required for the assay. Insert the strips in the frames for use. Re-bag any unused strips and frames, • Avoid the use of hemolyzed or lipemic sera. and store desiccated at 2°C to 8°C for future use. The silica pack in the bag keeps the plate dry, and turns from blue to pink if the bag is not properly sealed. • If large amounts of particulate matter are present in the sample, centrifuge or filter sample prior to analysis. Bind antigen Prepare samples a. Add 50 μL of standards or samples (see “Prepare samples” on page 2) to the appropriate wells. Sample concentrations should be within the range of the standard curve. Because conditions may vary, each investigator should determine b. Add 75 μL of 1X Assay Buffer into wells for detecting non-specific binding (NSB). the optimal dilution for each application. c. Add 50 μL of 1X Assay Buffer into maximum binding wells (B0 or Zero standard). Use all samples within 2 hours of dilution, or store at –20°C or lower until ready to perform assay. d. Add 25 μL of Allopregnanolone Conjugate to each well. Sample type Procedure e. Add 25 μL of Allopregnanolone Antibody to each well except NSB wells. f. Tap the side of the plate to mix. Cover the plate with plate sealer. Serum and plasma 1. Add diethyl ether or ethyl acetate to samples at a 5:1 (v/v) solvent:sample ratio. g. Shake at room temperature for 2 hours. If the plate is not shaken signals bound will be ~ 45% lower. 2. Mix solutions by vortexing for 2 minutes. Allow solvent layer to separate for 5 minutes. OR 3. Freeze samples in a dry ice/ethanol bath and pour solvent solution from the top of the sample into a Shake the plate in a plate shaker at room temperature for 15 minutes to ensure adequate mixing of the clean tube. Repeat steps 1-3 for maximum extraction efficiency, combining top layer of ether solutions. reagents. Incubate at 4°C for 16-18 hours. 4. Dry pooled solvent samples down in a Speedvac for 2-3 hrs, or under a nitrogen stream until dry. If h. Thoroughly aspirate the solution and wash wells 4 times with 300 μL of 1X Wash Buffer. samples need to be stored they should be kept desiccated at -20°C. Add chromogen 5. Redissolve samples at room temperature in 1X Assay Buffer. Use a minimum of 125 μL of Assay Buffer. a. Add 100 μL TMB Substrate to each well. The substrate solution will begin to turn blue. Urine Dilute samples ≥1:4 with 1X Assay Buffer. b. Incubate for 30 minutes at room temperature without shaking. Note: A Urinary Creatinine Detection Kit (Cat. no. EIACUN) is available for measuring urine creatinine for Note: TMB should not touch aluminum foil or other metals. normalization of allopregnanolone in a random urine specimen. Add stop solution Dried feces See detailed extraction protocol on the product page at thermofisher.com Add 50 μL Stop Solution to each well. Tap side of the plate gently to mix. The solution in the wells changes from Note: The ethanol concentration in the final diluted Assay Buffer dilution added to the well should be <5%. blue to yellow. Tissue culture media Dilute sample with 1X Assay Buffer. The investigator should determine the appropriate dilution for media samples prior to running the assay.

Dilute standards Note: Use glass or plastic tubes for diluting standards. Read the plate and generate the standard curve The Allopregnanolone Standard contains an organic solvent. Pipette the standard up and down several times to wet the pipet tip before 1. Read the absorbance at 450 nm. Read the plate within 10 minutes after adding the Stop Solution. transfer to ensure that volumes are accurate. 2. Average the duplicate Optical Density (OD) values for each standard and sample. Use curve-fitting software to generate the standard curve. A four parameter algorithm provides the best standard curve fit. Optimally, the background absorbance may be subtracted 1. Add 50 µL Allopregnanolone Standard to one tube containing 450 µL 1X Assay Buffer and label as 10,000 pg/mL allopregnanolone. from all data points, including standards, unknowns and controls, prior to plotting. 2. Add 250 µL Standard Diluent Buffer to each of 7 tubes labeled as follows: 5000, 2500, 1250, 625, 312.5, 156.25, and 0 pg/mL 3. Calculate the concentrations for unknown samples and controls from the %B/B0 curve. Multiply value(s) obtained for sample(s) by allopregnanolone. the appropriate factor to correct for the sample dilution. 3. Make serial dilutions of the standard as described below in the dilution diagram. Mix thoroughly between steps. Note: Dilute samples producing signals lower than that of the highest standard in 1X Assay Buffer and reanalyze. Multiply the 4. Use the standards within 2 hours of preparation. concentration by the appropriate dilution factor.

Performance characteristics Intra-assay precision Standard curve (example) Samples were assayed in replicates of 20 to determine precision within an assay. The following data were obtained for the various standards over the range of 0–10,000 pg/mL allopregnanolone. Parameters Sample 1 Sample 2 Sample 3 Note: 1 pg/mL of allopregnanolone is equivalent to 3.14 pM. Mean (pg/mL) 3,576.5 1,768.8 545.7 Std Allopregnanolone (pg/mL) Net Optical Density (450 nm) %B/B0 %CV 3.7 4.6 12.9 10,000 0.118 12.37 CV = Coefficient of Variation 5,000 0.229 24.00 Inter-assay precision 2,500 0.396 41.51 Samples were assayed in duplicates in 20 assay runs by four 1,250 0.583 61.11 operators to determine precision between assays. 625 0.721 75.58 Parameters Sample 1 Sample 2 Sample 3 312.5 0.849 88.99 Mean (pg/mL) 3,538.8 1,710.6 499.2 156.25 0.885 92.77 %CV 6.2 7.0 11.2 0 0.954 100 CV = Coefficient of Variation

Note: The NSB gave a mean OD value of 0.063.

2 ELISA Kit Product Information Sheet ELISA Kit Product Information Sheet 3 Performance characteristics, continued

Expected values Urine Allopregnanolone Competitive ELISA Kit High Low A number of diethyl ether-extracted serum samples from pregnant Expected Conc. Observed Conc. % Catalog Number EIAALLO (96 tests) Sample Sample humans were tested in the assay. Adjusted neat concentrations of (pg/mL) (pg/mL) % % Recovery Pub. No. MAN0018776 Rev B.0

allopregnanolone was >2,900 pg/mL. A number of serum samples from non-pregnant human samples were extracted and tested in 80 20 3,441.7 3,493.3 101.5 Note: For safety and biohazard guidelines, see the “Safety” appendix in the ELISA Technical Guide (Pub. no. MAN0006706). Read the Safety the assay. Adjusted neat concentrations of allopregnanolone 60 40 2,691.7 2,811.0 104.4 Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. ranged from 738 to >1,698 pg/mL. Urine samples, diluted 1:4 to 40 60 1,941.7 1,860.0 95.8 1:16 from pregnant and non-pregnant human and other Product description 20 80 1,191.6 1,078.3 90.5 mammalian species were tested in the assay. Adjusted The Allopregnanolone ELISA Kit is a solid-phase monoclonal antibody based competitive Enzyme-Linked Immunosorbent Assay (ELISA). concentration of allopregnanolone varied from 1,500 to >46,000 Mean Recovery 98.1% This assay is designed to detect and quantify the level of allopregnanolone in extracted serum, plasma, and dried fecal samples, or urine pg/mL for non-pregnant and 7,700 to >57,000 pg/mL for pregnant and tissue culture media. The assay recognizes allopregnanolone independent of species. samples. Specificity The following cross reactants were tested in the assay and Sensitivity calculated at the 50% binding point. Contents and storage The analytical sensitivity of allopregnanolone is 129.7 pg/mL. This Kit and components are shipped at –20°C. Upon receipt, store the kit at –20°C. Once open, store the kit at 4°C and use within 2 weeks. Steroid Cross-reactivity % was determined by adding two standard deviations to the mean Store the Allopregnanolone Conjugate at –20°C or lower after opening. O.D. obtained when the zero and Std7 were assayed 19 times, and Allopregnanolone 100 calculating the corresponding concentration. Pregnanolone 2.19 Components Quantity

Linearity Tetrahydrodeoxycorticosterone (THDOC) 3.08 Allopregnanolone Standard; 100,000 pg/mL allopregnanolone 125 µL Linearity was determined for fecal extracts and diluted urine by Dihydrodeoxycorticosterone (DHDOC) <0.08 Assay Buffer Concentrate (5X) 28 mL taking two samples, one with a low level and one with a higher Dihydrotestosterone 0.095 level of allopregnanolone, and mixing them in the ratios given Antibody Coated Wells, 96-well strip-well plate coated with goat anti-mouse IgG 1 plate Tetrahydrocorticosterone <0.08 below. The measured concentrations were compared to the Allopregnanolone Antibody 3 mL expected values based on the ratios used. 5α-dihydroprogesterone <0.08 Fecal Extract Corticosterone <0.08 Allopregnanolone Conjugate 3 mL High Low Estrone <0.08 Expected Conc. Observed Conc. % Wash Buffer Concentrate (20X) 30 mL Sample Sample (pg/mL) (pg/mL) Recovery Progesterone 0.12 % % TMB (Tetramethylbenzidine) Substrate 11 mL 11α-hydroxyprogesterone <0.08 80 20 2,046.9 2,027.5 99.1 20α-hydroxyprogesterone <0.08 Stop Solution; contains 1 M HCl, CAUSTIC 5 mL 60 40 1,625.1 1,580.4 97.2 Cortisone <0.08 Plate Sealer 1 40 60 1,203.4 1,187.6 98.7 Cortisol <0.08 20 80 781.7 764.3 97.8 Estradiol <0.08 Materials required but not supplied Prepare 1X Wash Buffer Mean Recovery 98.2% 1. Dilute 15 mL of Wash Solution Concentrate (20X) with Testosterone <0.08 • Distilled or deionized water 285 mL of deionized or distilled water. Label as 1X Wash • Microtiter plate reader with software capable of measurement at or near 450 nm (preferably with correction between 570 nm Buffer.

and 590 nm). 2. Store the concentrate and 1X Wash Buffer in the refrigerator. Limited product warranty Use the diluted buffer within 3 months. • Plate washer–automated or manual (squirt bottle, manifold Life Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies' General Terms and dispenser, or equivalent) Conditions of Sale found on Life Technologies' website at www.thermofisher.com/us/en/home/global/terms -and-conditions.html . If Prepare 1X Assay Buffer • Calibrated adjustable precision pipettes and glass or plastic you have any questions, please contact Life Technologies at www.thermofisher.com/support. 1. Dilute 14 mL of Assay Buffer (5X) with 56 mL of deionized or tubes for diluting solution distilled water. Label as 1X Assay Buffer.

Consult 2. Store the concentrate and 1X Assay Buffer in the refrigerator. Catalog Batch Temperature Caution, consult Procedural guidelines Use by Manufacturer instructions for accompanying documents 1X Assay Buffer is stable at 4°C for 3 months. Number code limitation use IMPORTANT! Reagents are lot-specific. Do not mix or Manufacturer: Life Technologies Corporation | 7335 Executive Way | Frederick, MD 21704 | USA interchange different reagent lots from various kit lots. The information in this guide is subject to change without notice. • Review the Procedural guidelines and Plate washing

DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, directions in the ELISA Technical Guide available at MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT. thermofisher.com. Important Licensing Information: This product may be covered by one or more Limited Use Label Licenses. By use of this product, you accept the terms and conditions of all applicable Limited Use Label Licenses. • Allow reagents to reach room temperature before use. Mix to © 2020 Thermo Fisher Scientific Inc. All rights reserved. MAGPIX, FLEXMAP 3D, xPONENT, Luminex, Luminex 100, and Luminex 200 are trademarks of Luminex Corporation. Excel redissolve any precipitated salts. is a trademark of Microsoft Corporation. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. • Solutions containing sodium azide will inhibit the activity of the peroxidase conjugate. Ensure that there is no contamination of labware or the plate washer with azide containing solutions.

thermofisher.com/support | thermofisher.com/askaquestion For research use only. Not for use in diagnostic procedures. thermofisher.com 01 October 2020