Overview: The new ProteoExtract® Protein Precipitation Kit provides efficient concentration of proteins and removal of interfering substances from dilute protein samples in a single step. The ProteoExtract Protein Precipitation Kit uses a unique combination of precipitation agents to quantitatively precipitate proteins and remove interfering substances, for one-step concentration and clean-up of proteins.
The ProteoExtract Protein Precipitation Kit delivers protein solutions with very low conductivity, making it ideally suited for a wide range of proteomics applications including IEF, 1DGE, 2DGE, and MS.
Features: - Highest yields and quantitative protein recovery including low-molecular weight proteins - High solubility of protein pellets
Fig.1: Protein methods by various methods Protein at a concentration of 2 mg/ml was precipitated using the ProteoExtract® Kit; protein pellets were redissolved in IEF buffer prior to determination of protein concentration. The same experiment was performed in parallel using TCA/deoxycholate (DOC) and products from Supplier I and Supplier II. All experiments were performed in duplicate. The ProteoExtract Protein Precipitation Kit delivers up to 3 times greater protein yields as compared to other precipitation methods.
Fig.2: Quantitative recovery of proteins Equal volume equivalents from samples precipitated with different methods were separated by 1D SDS-PAGE and visualized by Coomassie™ blue staining. Protein patterns from the ProteoExtract-precipitated proteins (P) are identical to the pattern from the liver extract (L), clearly demonstrating the quantitative recovery of proteins. Weaker staining is achieved by TCA/DOC indicating protein loss. Selective protein loss is observed using products from Supplier I (SI) and Supplier II (SII). M: molecular weight markers.
Roll-over a protein identity on the list to locate it on the gel
Spot
Identity
1
complement factor B Precursor
2
Gelsolin precursor
3
human serum albumin
4
transferrin n-terminal lobe
5
antithrombin III, chain L
6
haptoglobin chain b
7
transthyretin
8
inter-a trypsin inhibitor - related protein precursor
9
transferrin fragment HUMTF12 NID
10
apolipoprotein L1 precursor
11
a-1-antitrypsin chain A
12
haptoglobin chain a 2
Fig. 3: The ProteoExtract® Protein Precipitation kit is fully compatible with 2DGE and mass spectrometry. Human serum was subjected to albumin/IgG removal using the ProteoExtract® Removal Kit (Cat. No. 122642). 1 mg protein was precipitated with the ProteoExtract® Precipitation Kit, resolubilized in IEF buffer and subjected to 2DGE. Selected spots were excised and proteins digested using the ProteoExtract® All-in-One Trypsin Digestion Kit (Cat. No. 650212). Protein identities were obtained with high statistical significance demonstrating that the ProteoExtract® Protein Precipitation Kit does not introduce any protein modifications that might interfere with mass spectrometry.
For additional product information, insert, MSDS, and C of A, please visit the product detail page for the ProteoExtract® Protein Precipitation Kit, Cat. No. 539180
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For additional product information, insert, MSDS, and C of A, please visit the product detail page for the ProteoExtract
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