siRNA / RNAi /miRNA transfection Human Cal 27 cells

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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.

Proteins Protein quantification Fluorimetric method

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction

DNA PCR Hot start PCR Mammalian DNA

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

DNA PCR Methylation specific PCR Bacterial DNA

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

DNA PCR Methylation specific PCR Mammalian DNA

I am currently using a recombinant protein which shows metal-dependent DNase activity. Is it possible to pinpoint the source of the DNase activity after protein purification? More specifically, can I ensure that the DNase activity is not because of nuclease contamination from the E.coli that might have persisted and passed with the protein of interest during purification?

Discussions Is a bacterial nuclease contamination possible during protein purification?

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality hot-start DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction

DNA PCR Hot start PCR Bacterial DNA

Get tips on using COL1A Antibody (COL-1): sc-59772 to perform Western blotting Type I collagen

Products Santa Cruz Biotechnology COL1A Antibody (COL-1): sc-59772

Get tips on using Anti-Collagen I antibody [COL-1] (ab90395) to perform Western blotting Type I collagen

Products Abcam Anti-Collagen I antibody [COL-1] (ab90395)

Get tips on using Collagen I antibody to perform Western blotting Type I collagen

Products GeneTex Collagen I antibody

Get tips on using Anti-Collagen X antibody (ab58632) to perform Immunohistochemistry Mouse - Col X

Products Abcam Anti-Collagen X antibody (ab58632)

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