Protein expression and purification Insect cells Hi5

Get tips on using pET-28a(+)-g-α13 to perform Protein Expression Prokaryotic cells - E. coli α-13 giardin

Get tips on using pET20b-chIL-7/H to perform Protein Expression Prokaryotic cells - E. coli chicken IL-7

Get tips on using pFBDM-PS1(wt)-γ-secretase to perform Protein Expression Eukaryotic cells - S. frugiperda γ-secretase

Get tips on using pGAPZA-α-MF-op-rLlAlp2 to perform Protein Expression Eukaryotic cells - P. pastoris Alkaline phytase

Get tips on using pTRAkc-rbcs1-cTP/pRIC 3.0 to perform Protein Expression Prokaryotic cells - A. tumefaciens BFDV cp

Get tips on using pIEX-4-Crypα-6His-mMBP-UMODpXR to perform Protein Expression Eukaryotic cells - S. frugiperda mMBP

Get tips on using NE-PER™ Nuclear and Cytoplasmic Extraction Reagents to perform Protein isolation Mammalian cells - HLE-B3

The most widely used method for protein quantification is by spectrophotometry. The concentration of the protein in the samples is measured at an absorbance of 280 nm. The absorbance of the sample protein is then plotted against a standard curve. This method allows for total protein quantification in a sample (cell and tissue extracts). Before analysing the concentration of protein in the sample, it is important to choose the right test method.  For high protein concentration samples (above 5 - 160 mg/ml) the best method is to use the Biuret test. For low concentrations samples (between 1 - 2000µg/ml) the best methods are Lowry assay, BCA assay, Bradford assay and coomassie blue (for exact sensitivity of the test kits you use, refer to manufacturer's protocol). If the samples contain detergents like Triton X-100 then BCA assay is the best choice. For samples that have proteins larger than 3 KDa in size Bradford assay is the best choice. Each method has advantages and disadvantages, plan your analysis considering your sample characteristics.

The most widely used method for protein quantification is by spectrophotometry. The concentration of the protein in the samples is measured at an absorbance of 280 nm. The absorbance of the sample protein is then plotted against a standard curve. This method allows for total protein quantification in a sample (cell and tissue extracts). Before analysing the concentration of protein in the sample, it is important to choose the right test method.  For high protein concentration samples (above 5 - 160 mg/ml) the best method is to use the Biuret test. For low concentrations samples (between 1 - 2000µg/ml) the best methods are Lowry assay, BCA assay, Bradford assay and coomassie blue (for exact sensitivity of the test kits you use, refer to manufacturer's protocol). If the samples contain detergents like Triton X-100 then BCA assay is the best choice. For samples that have proteins larger than 3 KDa in size Bradford assay is the best choice. Each method has advantages and disadvantages, plan your analysis considering your sample characteristics.

Get tips on using pST44-yAda3Δ2HIS-yAda2Δ1-yGcn5 to perform Protein Expression Prokaryotic cells - E. coli Ada2/Ada3/Gcn5 complex