When looking for the activation of cell signaling pathways in the research domain, a phosphorylation study is important in getting the right results when the phospho-protein analysis is being carried out.
Phosphorylation entails adding phosphate groups to protein. It is a crucial, reversible post-translational protein modification. This is the mechanism meant for regulating the functional properties of proteins involved in gene expression, cell adhesion, cell proliferation, cell differentiation, and cell cycle. In studying the function of a specific protein, the determination of the Phosphorylation state is essential as it regulates the function and drives cellular signaling pathways.
What is Phospho-specific Antibodies?
Phospho-specific antibodies are affinity-purified and can be polyclonal or monoclonal antibodies. Phospho-specific antibodies can discriminate between the phosphorylated and non-phosphorylated forms of protein. Phospho-specific antibodies detect only the phosphorylated protein forms in a complex protein mixture in the cells. They can detect phosphorylated amino acids within the context of other surrounding amino acids in the sequence. This is done through the use of pan-antibody and phospho-specific antibodies. It continues to show essential roles that the tight regulations of phosphorylation proteins statuses are key for homeostasis and integral cellular processes. And this involves cell growth, cell cycle, apoptosis, and other cell death processes.
Why Phospho-specific antibodies are used?
Most researchers use phospho-specific antibodies because the status of individual phosphorylation sites is allowed to be determined easily. And with this, the active state of the protein indication is provided. Phospho-specific antibodies are used in various immunodetection applications such as the western blot, immunohistochemistry, and flow cytometry.
Seven popular photo-specific antibodies
Here are selections of some of the most popular phospho-specific antibodies that researchers use in carrying out their analysis.
- STAT 3 antibody
- AKT (Phospho-Ser 1473) antibody
- ATM (Phospho-Ser 1981) antibody
- SMAD2 (Phospho-Ser467) antibody
- BRCA1 (Phospho-Ser1524) antibody
- ISLET1 Antibody
- ERK1/2(PhosphoThr202/Tyr204) antibody
STAT3 Antibody: signal transducers and activators of transcription 3 (STAT 3) are a family of STAT proteins phosphorylated through receptors associated with kinases and translocate to the nucleus. In response to cytokines and growth factors, it acts as transcription components. STAT3 Antibody can be activated by Interferon-alpha, Interferon-gamma, EGF, PDGF, and IL6. STAT3 is responsible for mediating the expression of different genes, responding to cell stimuli. With this, it plays a vital part in many cellular processes like cell growth and apoptosis.
AKT (Phospho-ser 473) Antibody is also known as protein kinase B (PKB) or Rac. In controlling survival and apoptosis, AKT plays a critical role. It becomes activated following its phosphorylation at two regulatory residues, phospholipid binding and phosphorylation within the carboxy terminus at Ser473. The Antibody promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets.
ATM (Phospho-Ser 1981) Antibody: Ataxia-telangiectasia Mutated (ATM) is a protein belonging to the P13/P14 Kinase family. It regulates the cell cycle checkpoints and repair of DNA. The ATM gene codes for a protein belonging to the phosphoinositide -3 kinase superfamily. It becomes autophosphorylated and unregulated by exposure to ionizing radiation. This Antibody repairs double-stranded DNA breaks that occur because of ionizing radiation and other mutagens.
SMAD2 (Phospho-Ser467) antibody: SMAD2 Antibody regulates multiple cellular processes like cell proliferation, differentiation, and apoptosis. Its components belong to the critical intracellular pathways that transmit TGF-β signals from the cell surface into the nucleus. SMAD 2 proteins are the mammalian homologs of Drosophila mothers against decapentaplegic (Mad). It is implicated as the downstream effector of TGF-β/BMP signaling.
BRCA1 Antibody: This Antibody is responsible for detecting endogenous levels of total BRCA1 protein (breast and ovarian cancer susceptible). It is a high monoclonal antibody and is available both as the non-conjugated anti-BRCA1 antibody form and multiple conjugated anti-BRCA1. It includes agarose, HRP, PE, FITC, and multiple Alexa Fluor conjugates.
ISLET1 Antibody: it is a high-quality monoclonal islet-1 antibody suitable for detecting an islet-1 protein of rat, mouse, and human origins. This Antibody is available both as non-conjugated anti-Islet-1 and multiple conjugated anti-islet-1 antibody forms. It includes agarose, HRP, PE, FITC, and multiple Alexa Fluor® conjugates. Islet1 is in the family of LIM homeodomain transcription factors binding to the enhancer region of the insulin gene. In the regulations of insulin gene expression, it plays a vital role.
ERK1/2(PhosphoThr202/Tyr204) antibody: ERK1/2 belongs to the family of mitogen-activated kinases (MAPKs) of serine and threonine protein kinases. In regulating meiosis, mitosis, and postmitotic functions in cells, ERK1/2 are involved. They are activated by various extracellular stimuli like mitogen, growth factors, neurotransmitters, chemokines, and cytokines. The activation makes use of receptor tyrosine kinases (RTK), G protein-coupled receptors (GPCRs), or protein kinase C (PKC). After it has been stimulated, ERK1/2 becomes phosphorylated through the upstream kinases MEK on residues of Thr202/Tyr204.