What is a phospho-specific antibody?
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Phosphospecific antibodies are affinity-purified rabbit polyclonal or monoclonal antibodies that are monospecific for a target protein that is phosphorylated. These antibodies provide critical insight into the investigation of complex signal transductions events.
How are phospho antibodies made?
Phospho-specific antibodies are generated using peptides containing one or more phosphorylated amino acids. There are three residues which can be phosphorylated: Serine (S), Threonine (T) and Tyrosine (Y).
How do you determine if a protein is a phosphoprotein?
Scientists usually detect phosphoproteins by labeling them with radioactive phosphorous or by using antibodies that recognize specific phosphorylated amino acid residues.
How do you phosphorylate proteins?
Protein phosphorylation is a reversible post-translational modification of proteins in which an amino acid residue is phosphorylated by a protein kinase by the addition of a covalently bound phosphate group.
How do you detect hyperphosphorylation?
Methods for Detecting Protein Phosphorylation
- Introduction.
- Kinase Activity Assays.
- Phospho-Specific Antibody Development.
- Western Blot.
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Cell-Based ELISA.
- Intracellular Flow Cytometry and ICC/IHC.
- Mass Spectrometry.
Which is the example of phospho protein *?
Phosphoproteins are conjugate proteins, post-translationally modified by the attachment of phosphate group. Common examples of naturally occurring phosphoproteins are milk protein casein and ovo-vitellin of egg yolk.
What does a phosphoprotein do?
MYC is a nuclear phosphoprotein that functions as a sequence-specific DNA-binding transcriptional regulator controlling the transcription of target genes with diverse roles in regulating cell growth by affecting DNA replication, energy metabolism, protein synthesis, and telomere elongation [139–141].
Do kinases phosphorylate lipids?
Lipid kinases. Lipid kinases phosphorylate lipids in the cell, both on the plasma membrane as well as on the membranes of the organelles. The addition of phosphate groups can change the reactivity and localization of the lipid and can be used in signal transmission.
Do kinases phosphorylate?
For protein targets, kinases can phosphorylate the amino acids serine, threonine, and tyrosine.
How do you calculate kinase activity?
Kinase activity within a biological sample is commonly measured in vitro by incubating the immunoprecipitated kinase with an exogenous substrate in the presence of ATP.
How do you phosphorylate in vitro?
Phosphorylated proteins may be obtained 1) with in vitro phosphorylation by mixing a kinase with known activity with the protein-of-interest, or 2) by stimulating cells with known effect on the protein-of-interest. Unphosphorylated peptide or protein (negative control).