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NI™ (Non-Interfering™) Protein Assay

Catalog Description Size Price(USD) Qty
786-005
NI™ (Non-Interfering™) Protein Assay Kit with Albumin Standard 500 Assays
$274.00
786-896
NI™ (Non-Interfering™) Protein Assay Kit with Non Animal Protein Standard 500 Assays
$274.00

NI™ (Non-Interfering™) Protein Assay

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A highly sensitive, colorimetric protein assay that overcomes interference of common laboratory agents present in protein solutions and shows minimal protein-to-protein variation.

 
The assay is unaffected by the presence of common laboratory agents, such as reducing agents, chelating agents, detergents, amines, sugars, chaotropes, salts, drugs, antibiotics, cobalt and other common laboratory agents (see tables 1 and 2).
 
The NI™ Protein Assay is composed of two simple steps:

Universal Protein Precipitating Agent (UPPA™) is added to the protein solutions to rapidly precipitate total protein. Protein is immobilized by centrifugation and interfering agents in the supernatant are discarded.
Protein concentration is assayed by mixing with an alkaline solution containing a known concentration of copper salt; the copper ions bind to the peptide backbone and the assay measures the unbound copper ions (see figure 2). The assay is independent of protein side chains minimizing protein-to-protein variation (see figure 3). The color density is inversely proportional to the amount of protein.
 
The assay is supplied with a traditional bovine serum albumin (BSA) protein standard or a non animal protein standard.
 
Table 1: A selection of compounds and the maximum concentrations that are compatible with the NI™ Protein Assay.
Compound

Concentration

Compound

Concentration

Ammonium sulfate

40%

N-Octyl glucoside

0.5%

Brij® 35

1%

Phosphate buffer

0.2M

CHAPS

1%

Sarcosyl

1%

CHAPSO

1%

Sodium azide

0.1M

CTAB

1M

Sodium dodecyl sulfate (SDS)

1%

Digitonin

0.3%

Sucrose

30%

DTT

10mM

TCEP

15mM

EDTA

10mM

Thesit®

2%

Glycerol

30%

Thiourea

2M

Guanidine.HCl

6M

Tris

0.2M

Guanidine thiocyanate

6M

Triton® X-100

3%

HEPES

0.1M

Triton® X-114

1%

Iodoacetamide

15mM

Tween® 20

2%

2-mercaptoethanol

0.5%

Urea

8M

 
Table 2: NI™ Protein Assay is compatible with strong chaotropic extraction buffers
Buffer composition Buffer composition
4M urea, 1% SDS, 10mM EDTA, 0.8% 2-mercaptoethanol 1% Sarcosyl, 0.8% 2-mercaptoethanol, 4M guanidine thiocyanate, 10mM EDTA
6M urea, 2M thiourea, 4% CHAPS 6M urea, 2M thiourea, 2% CHAPS, 2% ND SB 201
6M urea, 2M thiourea, 4% Nonidet® P-40 6M urea, 2M thiourea, 2% CHAPS, 2% SB 2 10
 

Features
  • Linear response 0.5µg-50µg protein
  • Small sample requirement, only 1-50µl
  • Unaffected by non-protein chemicals and agents
  • Protocol time: ~30 minutes
  • Long Shelf Life

Applications

  • Estimate protein during protein purification, electrophoresis, cell biology, molecular biology, and other research applications.
  • Suitable for protein samples containing common laboratory agents, such as reducing agents
    (ß-mercaptoethanol, dithiothreitol), chelating agents (EDTA), detergents, amines (Tris), sugars and many other agents.
  • Suitable for samples containing chaotropic agents such as urea, thiourea, guanidine hydrochloride, guanidine thiocyanate, ammonium sulfate, drugs, antibiotics, cobalt, and numerous other agents and extraction buffers.
  • Suitable for determination of protein concentration in cellular fractions, tissue & cell lysates and chromatography purification fractions.
  • Suitable for dilute protein solutions.
Protocol
786-005
786-896
Material Safety Data Sheet
786-005
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786-896
English_US
Danish
Dutch
English_UK
French
German
Norwegian
Portuguese
Swedish
Polish
Technical Literature
A Non-Animal Alternative to BSA Protein Standards
Bioassays Handbook
Protein Assay Handbook & Selection Guide  An introduction to protein assays.
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  • Sumikawa, T. et al(2018) Site-Specific Integration by Recruitment of a Complex of ΦC31 Integrase and Donor DNA to a Target Site by Using a Tandem, Artificial Zinc-Finger Protein. Biochem. DOI: 10.1021/acs.biochem.8b00979.
  • Yumnam, S. et al (2018) Identification of a novel biomarker in tangeretin‑induced cell death in AGS human gastric cancer cells.Oncol. Rep. doi: 10.3892/or.2018.6730
  • Castano-Duque, L and Luthe D.S. (2017) Protein networks reveal organ-specific defense strategies in maize in response to an aboveground herbivore. Arthropod-Plant Interactions. https://doi.org/10.1007/s11829-017-9562-0
  • Taruno, A. et al (2017) Post-translational palmitoylation controls the voltage gating and lipid raft association of CALHM1 channel. J Physiol.DOI: 10.1113/JP274164
  • "Bennett, M. et al (2017) A Novel Biomarker Panel to Identify Steroid Resistance in Childhood Idiopathic Nephrotic Syndrome. Biomarker Insights. DOI: 10.1177/1177271917695832"
  • Nagappan, A. et al (2016) Proteomic analysis of selective cytotoxic anticancer properties of flavonoids isolated from Citrus platymamma on A549 human lung cancer cells Mol. Med. Rep. DOI: 10.3892/mmr.2016.5666
  • Zhou, Y. et al (2016) Lens ER-stress response during cataract development in Mip-mutant mice.Biochim. Biophys. Acta. doi:10.1016/j.bbadis.2016.05.003
  • Lohnes, K. L. et al (2016) Combining High-throughput MALDI-TOF Mass Spectrometry and Isoelectric Focusing Gel Electrophoresis for Virtual 2D Gel-based Proteomics. Methods. DOI:10.1016/j.ymeth.2016.01.013
  • Kaur, G. et al (2015) The peptidyl-prolyl cis-trans isomerase activity of the wheat cyclophilin, TaCypA-1, is essential for inducing thermotolerance in Escherichia coli. J. Biopen. 2:9
  • Kaur, G. et al (2015) PLOS. DOI: 10.1371/journal.pone.0136692
  • Ayyu A. et al (2015) Mol Med Rep. http://dx.doi.org/10.3892/mmr.2015.3822
  • Gorski, J.P. et al (2015) Deletion of Mbtps1 (PCSK8, S1P, SKI-1) in Osteocytes Stimulates Soleus Muscle Regeneration and Increased Size and Contractile Force with Age. Jbc. DOI:M115.686626
  • Vasani, R. B. et al (2015) Microwave Heating of Poly(N-isopropylacrylamide)-Conjugated Gold Nanoparticles for Temperature-Controlled Display of Concanavalin A. Appl. Mater. Interfaces DOI: 10.1021/acsami.5b08765
  • Wijeratne, A. B. et al (2015) ACS Appl Mater Interfaces. 7:11155
  • Sathish, S. et al (2015) POJ 8:201
  • Kim, J. A. et al (2014) BMC Complement Altern Med. 14:379
  • Rossi, O. et al (2014) Mol Biotechnol. 57:84
  • Castellanos-Mendoza, A. et al (2014) Microbial Cell Factories 13:137
  • Mi, Y. et al (2014) JBC. 289:12157-12167
  • McGuire, J.D. et al (2014) Bone. 63:29
  • Yang, X. et al (2014) J. Cell. Biochem. 115:141
  • Gallo, J. et al (2014) J. Mater. Chem. B. 2:868
  • Mi, Y. et al (2012) JBC. 289:12157
  • Liu, T. et al (2014) Int. J. Oncol. 44:467
  • McGuire, J.D. et al (2014) J. Dentistry. 42:626
  • Wang, Y. et al (2013) Tumor Biol. 34:3649
  • Blech-Hermoni, Y. et al (2013) Dev. Dynam. 242:767
  • Miner-Williams, W. et al (2013) J. Anim. Physiol. Anim. Nutr. 97:
  • Peters, N.T. et al (2013) Mol. Microbio. 89:690
  • Nagappan, A. et al (2013) BMC Biochem. 14:24
  • Liu, T. et al (2013) Int. J. Oncol. 43:1125
  • Dasgupta, T. et al (2013) PLoS ONE 8(2): e56590
  • Nagappan, A. et al (2013) Chem-Biol Interact. 206:143
  • Suliman, M. et al (2013) J. Proteomics. 78:508
  • Guillon, F. et al (2012) J Exp Bot 63:739
  • Miner-Williams, W. et al (2012) Am. J. Clin. Nutr. 96:508
  • Liu, T. et al (2012) Int. J. Oncol. 41:2087
  • Miner-Williams, W. et al (2012) Am. J. Clin. Nutr. 96:508
  • Mendias, C.L. et al (2012) Muscle Nerve. 45:55
  • del Castillo, C.S. et al (2012) Fish Shellfish Immun. 32:79
  • Guillon, F. et al (2011) J Exp Bot 10:1093
  • El-Osta, M.A. et al (2011) J Biol Chem 286:19340
  • Chang, L. et al (2011) Mol. Pharmaceutics. 8:1767
  • Sanchez, C.J. et al (2011) BMC Microbiol. 11:245
  • Park, S.B. et al (2011) PLoS ONE 6(3): e17629
  • Larre, C. et al (2010) J. Exp. Bot. 61:1771
  • Reddy, V.B. and Lerner, E.A. (2010) Brit. J. Dermatol. 163:532
  • Groves, B. et al (2010) PLoS ONE 5(1): e8877
  • Xu, Z. et al (2010) PLoS ONE 5(3): e9725
  • Xia, B. et al (2010) PLoS ONE 5(7): e11771
  • Karbarz, M. et al (2009) J Biol Chem 284:414
  • Eismann, T. et al (2009) Am J Physiol Gastr Liver Physiol 296:G266
  • Iibuchi, R. et al (2009) Jap. J. Vet. Res. 57:13
  • Xu, Z. et al (2009) J Cell Biol 186:343
  • Yan, W. et al (2009) Proteomics. 3:116
  • Miner-Williams, W. et al (2009) J. Agric. Food Chem. 57:2072
  • Rintala, E. et al (2009) BMC Genomics. 10:461
  • Haffey, W.D. et al (2009) Adv. Enzyme Regul. 49:121
  • Stella, C.L. et al (2009) Am. J. Obstet. Gynecol. 201:387.e1
  • Wong, J.H. et al (2009) J. Cereal Sci. 49:73
  • Urbonavicius, S. et al (2009) J. Vasc. Surg. 49:455
  • Bendezu, F. et al (2008) J Bacteriol 190:1792
  • Lin, F.L. et al (2008_ Polymer Preprints. 49:1095
  • Ramachandran, P. et al (2008) Clin. Proteomics. 4:80
  • Yu, T.Y. et al (2008) Biochemistry. 47:13942
  • Gorski, J.P. et al (2009) Cell Tissues Organs. 189:25
  • Salusjarvi, L. et al (2008) Microbial CEll Fact. 7:18
  • Kalsotra, A. et al (2008) PNAS 105:20333
  • Lyngholm, M. et al (2008) Exper. Eye Res. 87:96
  • Brobey, R.K. and Soong, L. (2007) Proteomics. 7:116
  • Devouge, V. et al (2007) J. Proteome Res. 6:1342
  • Vorum, H. et al (2007) Proteome Sci. 5:5
  • Shiels, A. et al (2007)Invest Ophthalmol Vis Sci. 48:500
  • Huffman, N. et al (2007) J. Biol. Chem. 282:26002
  • Werner, M. et al (2007) J Biol Chem 282:5560
  • Banowetz, G.M. et al (2007) J. Therm. Biol. 32:12
  • Donohoe, M.E. et al (2007) Molecular Cell. 25:43
  • Brobey, R.K. et al (2006) Braz. J. Infect. Dis. 10:1
  • Reddy, V. et al (2006) J Biol Chem 281:16197
  • Tao, T.Y. et al (2006) Plant Cell Rep. 25:848
  • DePinto, W. et al (2006) Mol Cancer Ther 5:2644
  • Lawrence, D. et al (2006) Glia. 53:81
  • Ramachandran, P. et al (2006) J. Proteome Res. 5:1493
  • Dennison, S. et al (2006) Biophysical J. 90:1661
  • Sun, B.K. et al (2006) Molec. Cell. 21:617
  • Nielsen, K. et al (2006) Exp. Eye Res. 82:201
  • Ho, T. et al (2005) Hum Mol Genet. 14:1539
  • Jarrold, B. et al (2005) Electrophoresis. 26:2269
  • Cao, Y.N. et al (2005) Biochem. Piophys. Res. Comm. 332:866
  • Park, G.S. et al (2005) Virology. 340:1
  • Sørensen, B.S. et al (2005) Radiother. Oncol. 76:187
  • Aoki, M. et al (2004) Acta Cryst. D60:439
  • Johnson, J. et al (2004) J Bacteriol 186:2481
  • Macvaughton, T.B. and Lai, M. (2004) Meth. Mol. Med. 95:99
  • Agnew, B.J. et al (2004) Electrophoresis. 25:2478
  • Wong, J. et al (2004) Plant Cell Physiol 45:407
  • Busenlehner, L.S. et al (2004) Biochemistry. 43:11145
  • Courville, T. et al (2004) J Biol Chem 279:3318
  • Banowetz, G.M. et al (2004) Anal Biochem. 332:314
  • Gushwa, N. et al (2003) Plant Physiol 132:1925
  • Lackner, L. et al (2003) J Bacteriol. 185: 735
  • Macnaughton, T. et al (2003) J Virol 77:12048
  • Clarke, B.L. (2003) Meth. Mol. Biol. 228:151
  • Saito, K. et al (2003) Molecular Microbiology. 48:373
  • Wu, K.H. et al (2003) Neuroendocrinol. 78:72
  • Loeb, D. et al (2003) Leukemia. 17:965
  • Endo, S. and Launey, T. (2003) Neuropharmacol. 45:863
  • Loeb, D. et al (2002) J Biol Chem 277:19627
  • Kilpatrick, L. et al (2002) Am J Physiol Cell Physiol 283:C48
  • Xu, N. et al (2002) Am J Physiol Lung Cell Mol Physiol 282:L796
  • Waskewich, C. et al (2002) Cancer Res. 62:2029
  • McClowry, T.L. et al (2002) J. Med. Virol. 66:96
  • Dennison, S.M. et al (2002) Biochemistry. 41:14925
  • Janabi, N. (2002) J. Immunol. 168:4747
  • Reeve, I. et al (2002) PNAS 99:8608
  • Sun, S. et al (2002) Anal. Biochem. 307:287
  • Blumenthal, E. et al (2001) Clin. Cancer. Res. 7:3178
  • Timmerman, M. M. and Woods, J. P (2001) Infect. Immun. 69:7671
  • Tikunov, B. et al (2001) J. Appl. Physiol. 90:1927
  • Kasahara, K. et al (2001) Clin Endocrinol Metab. 86:1281
  • Tatsumi, R. et al (2001) J. Musc. Res. Cell. Motil. 22:149
  • Ladd, A. et al (2001) Mol Cell Biol 21:1285
  • Poutanen, M. et al (Rap. Comm. Mass Spec. 15:1685
  • Vasara, T. et al (2001) Mol. Microbiol. 42:1349
  • Thibeault, D. et al (2001) J. Biol. Chem. 276:46678
  • Aoki, M. et al (2000) Acta Cryst. D56:1464
  • Konno, R. et al (2000) Arch. Toxicol. 74:473
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