Protein phosphorylation
[Protein phosphorylation]http://www.creative-proteomics.com/services/phosphorylation.htm is the most commonly studied area of post-translational modification since it plays a vital role in intracellular signal transduction and is involved in regulating cell cycle progression, differentiation, transformation, development, peptide hormone response, and adaptation. It has been estimated that one third of mammalian proteins may be phosphorylated and this modification often plays a key role in modulating protein function. Reversible protein phosphorylation, principally on serine, threonine or tyrosine residues, is one of the most important and well-studied post-translational modifications.
[protein n-acetylation]http://www.creative-proteomics.com/services/n-acetylation.htm, or the transfer of an acetyl group to nitrogen, occurs in almost all eukaryotic proteins through both irreversible and reversible mechanisms. N-terminal acetylation requires the cleavage of the N-terminal methionine by methionine aminopeptidase (MAP) before replacing the amino acid with an acetyl group from acetyl-CoA by N-acetyltransferase (NAT) enzymes. This type of acetylation is co-translational, in that N-terminus is acetylated on growing polypeptide chains that are still attached to the ribosome. While 80-90% of eukaryotic proteins are acetylated in this manner, the exact biological significance is still unclear.
[host cell protein hcp analysis]http://www.creative-proteomics.com/application/host-cell-protein-analysis.htm Because most of recombinant proteins are synthesized by cell-based systems in biochemical researches, the host cells derived from bacteria, yeast, mammalian cells, insect cells and plants (such as rice and tobacco) can be used for protein therapeutics manufacturing. But limited by current purification techniques, low levels (1 to 100 ppm) of host cell proteins (HCPs) may still remain in the purified biotherapeutics, even after a series of purifications. The ppm-level contaminants in biotherapeutics may trigger an unpredictable immune response in patients after dosing, and are required to be identified and quantified as part of drug safety evaluation, by the regulatory agencies.
The transfer of one-carbon methyl groups to nitrogen or oxygen (N- and O-methylation, respectively) to amino acid side chains increases the hydrophobicity of the protein and can neutralize a negative amino acid charge when bound to carboxylic acids. Methylation is mediated by [post translational modification methylation]http://www.creative-proteomics.com/services/methylation.htm, and S-adenosyl methionine (SAM) is the primary methyl group donor. Methylation occurs so often that SAM has been suggested to be the most-used substrate in enzymatic reactions afterATP. Additionally, while N-methylation is irreversible, O-methylation is potentially reversible.
Creative Biogene [Plant Direct PCR]http://www.creative-biogene.com/Plant-Direct-PCR-Kit-DPZ-08-1234416-87.html Kit is designed to amplify DNA directly from plant samples, such as such as leaf discs, seeds and crude plant extracts. The kit is based on a specially engineered DNA Polymerase with extremely robust and tolerant of many PCR inhibitors present in plant material. No DNA purification is required and unpurified DNA can be directly used as template for PCR.
Creative Biogene [Animal Tissue Direct PCR Kit]http://www.creative-biogene.com/Animal-Tissue-Direct-PCR-Kit-DPZ-03-1234411-87.html is designed for amplification of DNA from a wide variety of animal tissues including mice, fish, birds and insects. This kit contains a novel buffer system that effectively lyses tissue and neutralizes inhibitors. No DNA purification is required and unpurified DNA can be directly used as template for PCR.
[protein sequence analysis]http://www.creative-proteomics.com/services/de-novo-protein-sequence-analysis.htm (PMF) is an investigative method for protein distinguishing proof. Fundamentally, the obscure protein of intrigue is initially divided into littler peptides, whose outright masses can be precisely measured with a mass spectrometer, for example, MALDI-TOF or ESI-TOF. At that point these masses are contrasted with either a database containing known protein successions or even the genome grouping which can be converted into proteins through PC programs. At that point the total masses of the peptides from every protein are figured hypothetically for mass correlation between the peptides of the obscure protein and the hypothetical peptide masses of every protein to locate the best match.
The upside of PMF strategy is that lone the masses of the peptides is should be known, while tedious once more peptide sequencing is then pointless, the length of the protein arrangement is available in the database of intrigue. Furthermore, most PMF calculations accept that the peptides originate from a solitary protein while the nearness of a blend can altogether confound the investigation and conceivably trade off the outcomes, in this way a segregated protein is required for the PMF based protein distinguishing proof. Blends surpassing various 2-3 proteins commonly require the extra utilization of MS/MS based protein distinguishing proof to accomplish adequate specificity of recognizable proof.
Since a large portion of recombinant proteins are combined by [Host Cell Protein Analysis]http://www.creative-proteomics.com/application/host-cell-protein-analysis.htm frameworks in biochemical looks into, the host cells got from microorganisms, yeast, mammalian cells, bug cells and plants, (for example, rice and tobacco) can be utilized for protein therapeutics fabricating. Be that as it may, restricted by current cleaning procedures, low levels (1 to 100 ppm) of host cell proteins (HCPs) may at present stay in the purged biotherapeutics, even after a progression of decontaminations. The ppm-level contaminants in biotherapeutics may trigger an unusual resistant reaction in patients in the wake of dosing, and are required to be recognized and measured as a major aspect of medication wellbeing assessment, by the administrative offices.