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Nucleic Acid and Protein Quantitation Quant-iT™ Assay Kits for microplate-based assays of DNA, RNA, and protein Nucleic Acid and Protein Quantitation Quant-iT™ Assay Kits for microplate-based assays of DNA, RNA, and protein → Selective quantitation—more accurate than UV absorbance readings or Bradford assays → High sensitivity—use as little as 1 µl of your sample → Simple assay design—mix-and-read format for quantitation in microplates, with or without robotics → Convenient packaging—prediluted standards and buffers for streamlined quantitation A DNA + RNA (1:1) B RNA Quant-iT™ Assay Kits (Table 1) utilize 0 1.0 Fluorescence Better technology, better results Fluorescence DNA 2.0 DNA (ng) DNA advanced fluorophores that become fluorescent upon binding to DNA, RNA, RNA + DNA (1:1) RNA 0 20 60 40 80 100 20 0 40 DNA or RNA (ng) 60 80 100 RNA or DNA (ng) or proteins; the fluorescence intensity of the resulting complex depends directly C BSA Ovalbumin Lysozyme Pepsin IgG Histone B-Casein Myoglobin A-Casein in the sample. Quant-iT™ Technology gives more accurate results than can be obtained with UV absorbance read- Fluorescence on the amount of the target molecule ings or Bradford protein assays (Figure 1), because the Quant-iT™ Reagents only report the molecule of interest—there’s no interference from free nucleotides or other contaminants. And more accurate quantitation means higher confidence for the success of downstream experiments. 0 1.0 2.0 3.0 4.0 5.0 Protein (µg) Figure 1—Quantitation using Quant-iT™ Assay Kits. A The Quant-iT™ dsDNA Assay Kit, High Sensitivity has a linear detection range of 0.2–100 ng and is selective for dsDNA, even in the presence of an equal mass of RNA. The x-axis gives the mass of nucleic acid when DNA or RNA is assayed alone; in the 1:1 mixture, the total mass of nucleic acid is double the amount shown. The inset shows the sensitivity of the assay for DNA. B The Quant-iT™ RNA Assay Kit has a linear detection range of 5–100 ng and is selective for RNA, even in the presence of an equal mass of DNA. The x-axis gives the mass of nucleic acid when DNA or RNA is assayed alone; in the 1:1 mixture, the total mass of nucleic acid is double what is stated. C The Quant-iT™ Protein Assay Kit has a detection range of 0.25–5 μg. This graph shows the signal generated from nine different proteins over the stated detection range. The protein-to-protein variability at the 3 μg point for all nine proteins assayed is 12%. Save your samples It’s important to get accurate quantitation while still leaving sufficient sample for downstream applications. Quant-iT™ Reagents are up to 1,000 times more sensitive than UV absorbance readings; as little as 1 µl of your sample is all that's needed to get reliable results. The pro- Quant-iT™ Reagent tein assay is up to 20-fold more sensitive than standard Bradford assays. Quant-iT™ Assay Kits provide the ultimate in convenience for DNA, RNA, Working solution (1:200 dilution) and protein quantitation in a simple mixand-read microplate format (Figure 2). They are the best choice for quantitating Microplate Buffer 1. Load microplate with working solution 20–2,000 samples at a time. These Quant-iT™ Kits provide: → Concentrated Quant-iT™ Reagents → Prediluted standards for generating a standard curve → Ready-to-use 1X buffer → Protocols for high throughput and automation-level efficiency 2. Add standards and samples and mix well 3. Read plate Figure 2—DNA, RNA, and protein quantitation using the Quant-iT™ Assay Kits. The Quant-iT™ Assay Kits have been specially formulated with ready-to-use buffers, prediluted standards, and easy-to-follow instructions, making quantitation both accurate and extremely easy. www.invitrogen.com Table 1—Quant-iT™ Assay Kits. Product Features Concentration † Applications Quant-iT™ dsDNA Assay Kit, High Sensitivity, 1,000 assays *0.2–100 ng* Selective for low-concentration dsDNA; can be used to quantitate dsDNA in the presence of free nucleotides, RNA, and protein 10 pg/µl to 100 ng/µl dsDNA Genomic DNA Viral dsDNA PCR products Genotyping Quant-iT™ dsDNA Assay Kit, Broad Range, 1,000 assays *2–1,000 ng* Selective for high-concentration dsDNA; can be used to quantitate dsDNA in the presence of free nucleotides, RNA, and protein 100 ng/µl to 1 µg/µl dsDNA Plasmid preps Subcloning fragments DNA for transfection Quant-iT™ RNA Assay Kit, 1,000 assays *5–100 ng* Selective for RNA; can quantitate RNA in the presence of free nucleotides, DNA, and proteins 250 pg/µl to 100 ng/µl RNA Microarray samples Real-time PCR samples In vitro transcription Quant-iT™ Protein Assay Kit, 1,000 assays *0.25–5 µg* Selective for proteins; unaffected by DTT, β-mercapto ethanol, amino acids, and DNA contamination 12.5 µg/ml to 5 mg/ml protein PAGE gels Western blots Activity assays † Sample concentration assumes a 1–20 µl sample volume in a 200 µl assay. Ordering Information Product Quantity Cat. no. Quant-iT™ dsDNA Assay Kit, High Sensitivity 1 kit Q33120 Quant-iT™ dsDNA Assay Kit, Broad Range 1 kit Q33130 Quant-iT™ RNA Assay Kit 1 kit Q33140 Quant-iT™ Protein Assay Kit 1 kit Q33210 For more information, visit our website at probes.invitrogen.com/products/quantitation. www.invitrogen.com ©2006 Invitrogen Corporation. All rights reserved. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated. F-065430-r1 US 0506