Premiums products and value-added services V
Value-added products on the topic has continued for five, have you already had a general understanding of our value-added products? These series of topics’s writing are used to help our customers understand our product’s classification better, and find their own needs conveniently and accurately. Today we will continue to introduce three value-added products for everyone, followed by PCR Mix System, Lysis Buffer for Cells, Tissue, and Protein A/G Purification Column.
The first one is PCR Mix System.
PCR reagents are most widely used in bioscience research and almost all laboratories involved in molecular biology. At present, the Master Mix form of products is gradually becoming the mainstream of PCR reagent products as for its conveniences. Cloud-Clone Corp has many technical advantages of products, and we have established a strict production, quality control system, these can ensure that our products have a stable quality and a good user experience.
PCR Mix System provides 2 times of the concentration of PCR pre-mixed solution, together with Taq DNA polymerase, dNTPs, standard Taq enzyme reaction buffer, enzyme stabilizer and bromophenol blue dye, suitable for routine PCR. For the customer, it just needs to add templates and primers, PCR reaction could be quickly and easily completed, and the reaction products could be directly used for electrophoresis after PCR, so, it also reduces pollution and operating errors in the experiment.
1. Conventional PCR amplification; 2. DNA markers, such as isotopes and biotin labeling; DNA sequencing; 3. Gene scanning based on PCR; 4. T/A cloning for the 3'-terminal base A.
The next one is Lysis Buffer for Cells/Tissue;
Lysis Buffer is used to lyse cells for target protein extraction. The main functions are shown blew: 1. Use detergent to destroy lipid bilayer of cells; 2. Dissolve protein; 3. Denature protein and maintain its stability; 4. Inhibit protease activity.
Lysis buffer is used to lyse cells under non-denaturing buffer conditions, it consists of surfactant, a protease inhibitor, etc. It’s mainly used for protein extraction from cells and tissues. The four lysis buffers are optimized from traditional lysis buffer. They're suitable for lysis of cell membrane, cytoplasm and nucleus etc., which will help the release of proteins and do good for later extraction, ELISA, Western Blot, etc. We suggest 107 cells/50mg tissues by 1 ml of lysis buffer.
The types of lysates and their functions are shown below:
|Lysis Buffer 1||Membrane proteins including surface proteins, receptors|
|Lysis Buffer 2||Cytoplasmic proteins, structural proteins, etc.|
|Lysis Buffer 3||Cytoplasmic proteins, nuclear matrix proteins, membrane proteins, etc.|
|Lysis Buffer 4||Nuclear matrix proteins, endosome and other organelles protein|
The last one is Protein A/G Purification Column.
Recombinant protein A and CNBr activated beaded-form of agarose are coupled to make prepacked columns of protein A , it can specifically combined with Fc section of a variety of immunoglobulins.
Prepacked columns of protein A can be used for the purification of antibodies from different species and subtypes, and it could also be used for capturing immune complexes in Co-Immunoprecipitation. The product uses the multilocus crosslinking technology, it possesses good stability, high specificity and broad-spectrum binding force, 1 ml of the matrix can specifically combine with 5-10 mg of antibody. For example, antiserum not only contains a specific antibody against an antigen for immunization, but also have a large amount of hybrid proteins, polysaccharides and other blood components. In this case, the non-antibody component of the antiserum can be removed by using the Protein A /G purification column. Moreover, the antibody prepared directly from animal may appears high background and the gel suppressed for a long time will be blurred. Further purification is often required.
Through a lot of experimental data, we summarize some common problems in the process of protein A/G purification column, and give the corresponding solutions for each problem. Hope it is helpful to your use:
1. A common problem with Protein A or Protein G columns is that some antibodies do not bind well to the resin. In this case, it is recommended to increase the concentration of buffer salts used for binding and washing. The interaction of protein A or protein G with the Fc of the antibody is mainly due to hydrophobic interaction. Addition of high concentrations of salt contributes to their interaction and increases the binding force.
2. The antibodies may be destroyed with low pH. Antibodies of different species or subclasses should be eluted with different pH value of acidic solutions.The antibodies purified by the Protein A /G column could be eluted with the same pH value, except for a few cases, and even with some acidic eluents. The reagent with gradually decreased pH value could be used to confirm the most mild conditions to elute the intersted antibody.
3. Antibody purification columns may be used repeatedly. However, in analytical studies, please note the operation when we purify two or more antibodies by the same affinity column. As it is known, different antibodies have different affinity with protein A or protein G, and acid elution may not completely wash away the bounded immunoglobulins when they are closely fixed to the column, so, that would easily cause certain contamination by residual antibodies in the affinity column, and resulting in cross-reactivity. For large samples’ purification, it is recommended that a fixed affinity column should be used for antibodies from the same origin.
Today's presentation is coming to an end, and we will introduce a variety of customized services for everyone next time, please look forward to them!
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