Protein quantitation assay compatibility table

1 TECH TIP Protein assaysProtein quantitation assay compatibility tableNo single Protein assay is compatible with all sample components that might be encountered in a typical laboratory. To avoid the frustration of discovering incompatibility by trial and error, review the compatibility table on the following pages ( table 1) that summarizes compatible substances for several popular Protein assays. The table serves as a guide for assessing which Protein assay might be most effective with a given sample type. Use this information as a guide only. Since it is possible for sample and assay components to have an additive effect, we recommend verifying the compatibility of your specific sample buffer the compatibility tableThe table lists the maximum compatible concentrations for substances tested based on the midpoint concentration of a BSA standard. This is 1,000 g/mL for most assays and 10 g/mL for Thermo Scientific Pierce microplate BCA Protein assay kits. A substance was considered compatible at the indicated concentration if the error in Protein concentration estimation caused by the presence of the substance in a sample was less than or equal to 10%.

2 The Protein assays were conducted using the test tube procedure unless formulated for smaller volumes ( , Pierce microplate BCA Protein assays). The Pierce microplate BCA Protein assay kits and the Invitrogen Qubit Protein BR assay Kit were used with a microplate and PCR tube, and symbolsThe concentration listed refers to the actual concentration in the Protein sample. An denotes compounds that were not compatible at the lowest concentrations tested. NA indicates the substance has not been tested for that assay . Compounds are listed alphabetically under common names or abbreviations with the exception of sodium compounds, which are alphabetized under Na . Dilutions are expressed as undiluted or as a ratio, where 1:2 indicates 2-fold 1. Substances compatible with Thermo Scientific Pierce Protein assays and the Qubit Protein BR compound Pierce Rapid gold BCAP ierceBCA PierceMicroplate BCA-RAC* PierceMicro BCA PierceModified Lowry Pierce Detergent Compatible BradfordPierceCoomassie Plus Pierce Coomassie Pierce 660 nm Qubit Protein BR Assay2D sample buffer NA NA NA NA NA NANA NA Undiluted** NA2-mercaptoethanol 25 mM (35) 1 mM 1 mM 1 M1 M 1 M 1 M 1 mMACES, pH 25 mM 25 mM 10 mM NA NA100 mM 100 mM 50 mM NAAcetone 10% 10% 1% 10% 10%10% 10% 50% NAAcetonitrile 10% 10% 30% 1% 10% 10%10% 10% 50% 20%Ammonium sulfate 1 M1 M 1 M 125 mM 200 mMAprotinin 10 mg/L 10 mg/L 1 mg/L 10 mg/L 10 mg/mL10 mg/L 10 mg/L 2 mM NAAscorbic acid NA NA 1 mM 50 mM50 mM 50 mM 500 mM NAAsparagine NA 1 mM NA 5 mM NA10 mM 10 mM 40 mM NABicine 20 mM 20 mM 1 mM 2 mM NA 100 mM100 mM 100 mM >1 M 100 mMBis-Tris, pH NA33 mM mM mM NA 100 mM100 mM 100 mM 50 mM NABorate (50 mM)

3 , pH Undiluted Undiluted 1:4 NA UndilutedUndiluted Undiluted Undiluted UndilutedB-PER reagent Undiluted Undiluted 1:3 NA NA Undiluted1:2 1:2 1:2 UndilutedB-PER reagent II NA NA 1:4 NA NA NA1:4 NA 1:2 UndilutedB-PER reagent PBS NA NA 1:4 NA NA UndilutedNA NA 1:2 UndilutedBrij-35 5% 5% 5% 31% 1% 0 .125% 5% NABrij-56 NA 1% NA 1% 31% 31% NA NABrij-58 1% 1% 1% 1% 31% 5% NABromophenol blue (in 50 mM NaOH) NA 31% NACalcium chloride (in TBS, pH ) 10 mM 10 mM 1 mM 10 mM NA 10 mM10 mM 10 mM 40 mM NACesium bicarbonate NA100 mM 100 mM 50 mM NA100 mM 100 mM 100 mM NACetylpyridinium chloride NA NA NA NA NA NANA NA * * NACHAPS 5% 5% 10% (10) 1% 5%5% 5% 5% 5%CHAPSO 5% 5% 5% 31% 5%5% 5% 4% NACHES 100 mM 100 mM 50 mM 100 mM NA NA100 mM 100 mM >500 mM NACobalt chloride (in TBS, p H 7. 2 ) mM mM mM NA NA10 mM 10 mM 20 mM NACTAB NA NA NA NA NA NANA NA * * NACysteine NA mM 1 mM 10 mM10 mM 10 mM 350 mM NADithioerythritol (DTE) NA1 mM mM NA 1 mM 1 mM 25 mM NADithiothreitol (DTT) 1 mM 5 mM (5) 5 mM5 mM 5 mM 500 mM 5 mMDMF 10% 10% 5% 1% 10% 10%10% 10% 50% NADMSO 10% 10% 1% 10% 10%10% 10% 50% NADTAB NA NA NA NA NA NANA NA 2%* * 50 mMEDTA 10 mM 10 mM 5 mM (20) mM 1 mM 100 mM100 mM 100 mM 20 mM NAEGTA NA 5 mM (10) 1 mM 2 mM2 mM 2 mM 20 mM NAEPPS, pH 100 mM 100 mM 100 mM NA NA100 mM 100 mM 200 mM NAEthanol 10% 10% 1% 10% 10%10% 10% 50% NAFerric chloride (in TBS, p H 7.)

4 2 ) 10 mM 10 mM 5 mM mM NA NA10 mM 10 mM 5 mM NAGlucose 10 mM 10 mM 1 mM 100 mM NA1 mM 1 mM 500 mM 1 MGlutathione (reduced) NA NA 10 mM NA NA NANA NA 100 mM NAGlycerol (fresh) 10% 10% 5% 1% 10% 10%10% 10% 50% 10%Glycine-HCl, pH 100 mM 100 mM 50 mM NA 100 mM 100 mM100 mM 100 mM 100 mM Guanidine-HCl 4 M 4 M M (2) 4 M NA M M M 4 MHEPES, pH 100 mM 100 mM 200 mM (200) 100 mM 1 mM 100 mM100 mM 100 mM 100 mM NAHydrides (Na2BH4 or NaCNBH3) NA NA NA NANA NA NATest compound Pierce Rapid gold BCAP ierceBCA PierceMicroplate BCA-RAC* PierceMicro BCA PierceModified Lowry Pierce Detergent Compatible BradfordPierceCoomassie Plus Pierce Coomassie Pierce 660 nm Qubit Protein BR AssayHydrochloric acid (HCl) 100 mM 100 mM 10 mM 100 mM 100 mM100 mM 100 mM 125 mM NAImidazole, pH mM50 mM 30 mM (50) mM 25 mM 200 mM200 mM 200 mM 200 mM 200 mMI-PER reagent Undiluted Undiluted NA NA NA NANA NA 1:4 UndilutedLaemmli SDS sample buffer NA NALeupeptin 10 mg/L 10 mg/L 10 mg/L 10 mg/L 10 mg/L10 mg/L 10 mg/L 80 M NAMannitol NA NA NA NA NA NANA NA 100 mM NAMelibiose NA NA NA 25 mM NA100 mM 100 mM 500 mM NAMem-PER reagent Undiluted Undiluted 1:2 Undiluted NA NAUndiluted NA Undiluted UndilutedMem-PER Plus reagent UndilutedUndilutedNAUndilutedNAUndiluted NANAU ndilutedNAMES-buffered saline, pH Undiluted Undiluted 1:4 NA UndilutedUndiluted Undiluted Undiluted NAMES, pH 100 mM 100 mM 100 mM (100) 100 mM 125 mM 100 mM100 mM 100 mM 125 mM 125 mMMethanol NA10% 1% 10 % 10%10% 10% 50% NAMagnesium chloride NA NA 100 mM NA NA 500 mMNA NA >1 M NAModified Dulbecco s PBS NAUndiluted Undiluted Undiluted NA UndilutedUndiluted Undiluted Undiluted UndilutedMOPS, pH 100 mM 100 mM 200 mM 100 mM NA 100 mM100 mM 100 mM 125 mM 100 mMM-PER reagent Undiluted Undiluted 1:2 NA NA UndilutedUndiluted NA 1.

5 2 UndilutedN-acetylglucosamine 10 mM 10 mM NA NA100 mM 100 mM 100 mM NANa (sodium) acetate, pH 200 mM 200 mM 200 mM 200 mM 180 mM180 mM 180 mM 100 mM NANa azide 0 .125% NANa bicarbonate 100 mM 100 mM 100 mM 100 mM 100 mM100 mM 100 mM 100 mM NANa carbonate-bicarbonate, pH NAUndiluted Undiluted Undiluted NA UndilutedUndiluted Undiluted 1:3 MNa chloride NA1 M 150 mM 1 M 1 M 1 M5 M 5 M M 5 MNa citrate, pH 200 mM 200 mM 50 mM 5 mM NA 200 mM200 mM 200 mM mM NANa citrate-carbonate, pH 1:8 1:8 1:6 0 0 NA NAUndiluted Undiluted NANa citrate-MOPS, p H 7. 5 1:8 1:8 Undiluted 1:6 0 0 NA NANA Undiluted 1:16 NANa deoxycholate 5% 5% NA 5% NA 0 .1% NANa hydroxide (NaOH) 100 mM 100 mM 50 mM 100 mM 75 mM100 mM 100 mM 125 mM NANa phosphate 100 mM 100 mM 100 mM 100 mM 100 mM 100 mM100 mM 100 mM 500 mM NANE-PER reagent (CER) 1:2 Undiluted 1:2 NA NA Undiluted1:4 NA Undiluted UndilutedNE-PER reagent (NER) Undiluted Undiluted 1:4 NA NA NAUndiluted NA Undiluted UndilutedNickel chloride (in TBS, p H 7.)

6 2 ) NA10 mM mM NA NA10 mM 10 mM 10 mM NANP-40 5% 5% 5% 1% 5% 5%Octyl beta-glucoside 5% 5% (10) 0 .1% 31% 5% 5% NAOctylthioglucoside NA5% 7% 5% NA 5%3% 3% 10% NANa-orthovanadate (in PBS, pH ) 1 mM 1 mM mM 1 mM NA NA1 mM 1 mM 50 mM NAPhenol red NA g/mL NA mg/mL mg/mL mg/mL NAPhosphate-buffered saline (PBS) Undiluted Undiluted Undiluted Undiluted NA UndilutedUndiluted Undiluted Undiluted UndilutedPIPES pH NA100 mM 25 mM 100 mM NA NA100 mM 100 mM 100 mM NAPMSF in isopropanol 1 mM 1 mM 0 .125 m M 1 mM 1 mM 1 mM1 mM 1 mM 1 mM 1 mMPotassium thiocyanate 3 M 3 M NA 100 mM NA3 M 3 M 250 mM NAP-PER reagent NA 1:2 NA NA 1:2 NARIPA buffer Undiluted Undiluted 1:2 1:10 NA 1:41:40 1:10 Undiluted UndilutedSDS 5% 5% 5% (10) 5% 1% 0 .125% , 5%* * 5%Test compound Pierce Rapid gold BCAP ierceBCA PierceMicroplate BCA-RAC* PierceMicro BCA PierceModified Lowry Pierce Detergent Compatible BradfordPierceCoomassie Plus Pierce Coomassie Pierce 660 nm Qubit Protein BR AssaySpan 20 1% NA 1% NA NASucrose 40% 40% 40% (40) 4% 7.

7 5 % 10%10% 10% 50% 20%TCEP NA NA 10 mM (10) NA NA 100 mMNA NA 40 mM NAThimerosal NAThiourea NA NA NA NA NA NANA NA 2 M NATLCK 0 .1 m g / L 0 .1 m g / L 0 .1 m g / L mg/L NA0 .1 m g /m L 0 .1 m g / L 5 mg/mL NATPCK 0 .1 m g / L 0 .1 m g / L 0 .1 m g / L 0 .1 m g / L NA0 .1 m g /m L 0 .1 m g / L 4 mg/mL NAT-PER reagent 1:2 1:2 NA NA NA UndilutedUndiluted NA 1:2 UndilutedTricine, pH 25 mM 25 mM mM mM NA NA100 mM 100 mM 500 mM 50 mMTriethanolamine, pH 25 mM 25 mM 25 mM mM NA NA100 mM 100 mM 100 mM NATris-buffered saline (TBS) Undiluted Undiluted Undiluted 1:10 NA UndilutedUndiluted Undiluted Undiluted UndilutedTris-glycine, pH 1:21:3 1:10 NA UndilutedUndiluted Undiluted Undiluted Tris-glycine-SDS, pH Undiluted Undiluted Undiluted NA Undiluted1:4 1:2 Undiluted** Tris-HCl, pH 250 mM 35 mM (50) 50 mM 10 mM 2 M2 M 2 M 250 mM 500 mMTris-HEPES-SDS 100 mMNA NA NA NA NANA NA Undiluted** UndilutedTr i to n X-10 0 5% 5% 7% (10) 5% 31% 1% 0.

8 125% 1% 5%Tr i t o n X-114 1% 1% 2% (2) 31% 1% 0 .125% NATriton X-305 1% 1% 1% 1% 31% NA0 .125% 9% NATriton X-405 1% 1% 1% 31% 5% NATween 20 5% 5% 10% (10) 5% 1% 31% 10% 3%Tween 60 5% 5% 5% NA 0 .1% 5% NATween 80 5% 5% 5% 31% 0 .1% 5% NAUrea 3 M 3 M 3 M (4) 3 M 3 M 3 M3 M 3 M 8 M 3 MY-PER reagent Undiluted NA NA NA NANA NA Y-PER Plus reagent Undiluted NA NA NA UndilutedUndiluted NA 1:2 NAZinc chloride (in TBS, p H 7. 2 ) 10 mM 10 mM mM NA NA10 mM 10 mM 10 mM NAZwittergent 3-14 NA 1% 2% (2) NA NA* Selected values for the regular Pierce BCA Protein assay Kit - Reducing Agent Compatible (RAC) are given in parentheses in the column for the Pierce Microplate BCA-RAC kit.** Value when the Pierce 660 nm assay is run using Thermo Scientific Ionic Detergent compatibility Reagent for Pierce 600 nm Protein assay (Cat. No. 22663). Compound (buffer) formulation is described more fully in the buffer formulations table ( table 2).

9 For Research Use Only. Not for use in diagnostic procedures. 2021 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. COL25278 0721 Find out more at 2. Buffer formulations used in compatibility Cat. No. 2D sample buffer 8 M urea, 4% CHAPS or 7 M urea, 2 M thiourea, 4% CHAPS Laemmli SDS sample buffer 65 mM Tris-HCl, 10% glycerol, 2% SDS, bromophenol blue LC2676 MES-buffered saline, pH M MES, 150 mM NaCl (pH ) 28390 Modified Dulbecco s PBS 8 mM sodium phosphate, 2 mM potassium phosphate, M NaCl, 10 mM KCl (pH ) 2 8 374 Na carbonate-bicarbonate, pH M sodium carbonate-bicarbonate (pH ) 28382 Na citrate-carbonate, pH M sodium citrate, M sodium-carbonate (pH ) Na citrate-MOPS, pH M sodium citrate, M MOPS (pH ) Phosphate-buffered saline (PBS) 100 mM sodium phosphate, 150 mM NaCl (pH ) 28372 RIPA buffer 50 mM Tris, 150 mM NaCl, sodium deoxycholate, 1% NP-40, SDS (pH ) 89900 Tris-buffered saline (TBS) 25 mM Tris, 150 mM NaCl (pH ) 28379 Tris-glycine, pH 25 mM Tris, 192 mM glycine (pH ) 28380 Tris-glycine-SDS, pH 25 mM Tris, 192 mM glycine, SDS (pH )

10 28378 Tris-HEPES-SDS 100 mM Tris, 100 mM HEPES, 3 mM SDS 28398 Ordering information QuantityCat. Rapid gold BCA Protein assay Kit250 mLA53226500 mLA53225 Pierce BCA Protein assay Kit500 mL232271 L23225 Pierce Microplate BCA Protein assay Kit - Reducing Agent Compatible275 mL23252 Pierce Micro BCA Protein assay Kit500 mL23235 Pierce Modified Lowry Protein assay Kit530 mL23240 Pierce Detergent Compatible Bradford assay Kit450 mL23246 Pierce Coomassie Plus (Bradford) assay Kit950 mL23236 Pierce Coomassie (Bradford) Protein assay Kit950 mL23200 Pierce 660 nm Protein assay Kit450 mL22662 Qubit Protein BR assay Kit100 assaysA50668500 assaysA50669