Antibodies specific to internal control protein from Cloud-Clone Corp. (II)
Internal control is a reference in the experiment system, regularly, internal controls in mammalian cell expression system are proteins expressed by house keeping genes. These proteins show relatively constant quantity in different tissue and cells. So, when Western Blot is designed for protein level test, the internal control protein could be considered as a reference to further ensure the accuracy of the result. Last time we introduced the three commonly used internal controls including beta-actin, beta-tubulin, GAPDH (glyceraldehyde-3-phosphate dehydrogenase). This time we’ll present other internal controls produced by Cloud-Clone Corp.
Sodium/potassium-transporting ATPase (Na+/K+-ATPase), is an integrated membrane protein. It is composed of 1023 amino acids with a molecular weight of 113KD. Na+/K+-ATPase is selected as a reference control when the expression of a membrane protein was detected by WB method. No longer worry about how to chose the internal control for membrane protein detection in WB test, it is recommended to try antibody specific to Na + / K + - ATPase PAG369Mi01 prepared by Cloud-clone Corp.
Cytochrome c oxidase IV (Cox IV) is a hetero oligomeric enzyme that usually contains 13 subunits, located in the mitochondrial inner membrane. It is an enzyme complex for mitochondrial respiratory chain. Given the Cox IV in the outer mitochondrial membrane and the plasma membrane, antibody against to Cox IV PAD286Hu01 is usually used to detect the Cox IV, a internal control for mitochondrial protein and for which the MW is about 16KD.
Proliferating Cell Nuclear Antigen (PCNA), is named because it only exists in normal proliferating cells and tumor cells.Moreover, the further research proves that PCNA is closely related to DNA synthesis and it plays an important role in the initiation of cell proliferation. PCNA also is a good indicator to reflect the cell proliferation. PCNA, synthesized and located in the nucleus, is composed of 261 amino acids with a molecular weight of 36KD. Cloud-Clone Corp. had prepared antibody specific to PCNA PAA591Mi01. The result of Western Blot shows that the antibody can recognize PCNA in some cell lysates( Fig. 1),. But for non-proliferating cells or cell treated with antiproliferation, PCNA is not a good choice.
Lamin B, also called nuclear lamina B, is the target of caspase family, the key factors of apoptosis. Lamin B is activated by caspase shearing and it is located in the nuclear envelope, which is composed of 586 amino acids with a molecular weight of 66KD. As lamin B is highly conserved among different species, it is often used as a internal control. When the target protein is a nuclear protein, the antibody specific to Lamin B PAF548Mi01 prepared by Cloud-Clone Corp. might be a good choice. But for the samples with no nuclear membrane, the Lamin B is not recommended. In addition, Lamin B is not suitable as a internal control for apoptosis.
Histone h3.1 is often cited as the most commonly used internal control for nucleus. As it is involved in the structure of chromatin in eukaryotic cells, its expression is relatively stable. Histone is combined with DNA and some other nucleic acid to form a nuclear protein, in other words, Histone is the main composition of the nuclear protein and it is very conservative. Its expression level will not be changed by treated or not. That is why it is selected as the internal control. Histone h3.1 is composed of 136 amino acids and its molecular weight is 15KD. Cloud-Clone Corp. had developed the antibody specific to Histone H3 PAA285Mi01, which can be used for detection of internal control for nucleus.
Moreover, some factors should be taken into account when choosing the suitable internal control to ensure the accuracy of experimental results, including the sample species origin, molecular weight of target protein, distribution and expression of the target protein and experimental environment.
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