Product Name :
Phospho-PKA C-alpha (PRKACA)-T197 polyclonal antibody Background :
This gene encodes one of the catalytic subunits of protein kinase A, which exists as a tetrameric holoenzyme with two regulatory subunits and two catalytic subunits, in its inactive form. cAMP causes the dissociation of the inactive holoenzyme into a dimer of regulatory subunits bound to four cAMP and two free monomeric catalytic subunits. Four different regulatory subunits and three catalytic subunits have been identified in humans. cAMP-dependent phosphorylation of proteins by protein kinase A is important to many cellular processes, including differentiation, proliferation, and apoptosis. Constitutive activation of this gene caused either by somatic mutations, or genomic duplications of regions that include this gene, have been associated with hyperplasias and adenomas of the adrenal cortex and are linked to corticotropin-independent Cushing's syndrome. Alternative splicing results in multiple transcript variants encoding different isoforms. Tissue-specific isoforms that differ at the N-terminus have been described, and these isoforms may differ in the post-translational modifications that occur at the N-terminus of some isoforms. Product :
1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2 Storage&Stability :
Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles. Specificity :
Phosphorylated Immunogen :
A synthetic phosphorylated peptide around T197 of human PKA C-alpha (PRKACA)(NP_002721.1). Conjugate :
Unconjugated Modification :
Phosphorylated

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  Western blot analysis of extracts of 293T cells, using Phospho-PKA C-alpha -T197 antibody at 1:1000 dilution. 293T cells were treated by Insulin for 10 minutes or treated by PMA/TPA for 30 minutes after serum-starvation overnight.
  Secondary antibody: HRP Goat Anti-Rabbit IgG at 1:10000 dilution.
  Lysates/proteins: 25ug per lane.
  Blocking buffer: 3% BSA.
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  Western blot analysis of extracts of various cell lines, using Phospho-PKA C-alpha -T197 pAb at 1:2000 dilution or PKA C-alpha antibody .HeLa cells were treated by Calyculin A at 37℃ for 30 minutes after serum-starvation overnight.C6 cells were treated by PMA/TPA at 37℃ for 30 minutes after serum-starvation overnight.
  Secondary antibody: HRP Goat Anti-Rabbit IgG at 1:10000 dilution.
  Lysates/proteins: 25ug per lane.
  Blocking buffer: 3% nonfat dry milk in TBST.
  Detection: ECL Basic Kit .
  Exposure time: 1s.

Bioworld Biotech only provide peptides for our antibodies and do not provide additional peptide customization services.

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Price/Size :

USD 368/1mg/vial

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Tips: 

For phospho antibody, we provide phospho peptide（0.5mg) and non-phospho peptide(0.5mg).

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Describe :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (WB) and Immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or IHC performed with the neutralized antibody.

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Formula:

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.

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Storage:

The freeze-dried powder is more stable. For short time at 2-8°C. For long term storage store at -20°C. 

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Note :

This product is for research use only (RUO only). Not for use in diagnostic or therapeutic procedures.

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