Western blot analysis of human protein kinase A catalytic (PKAc) alpha subunit was performed by loading 40 µg of Jlat-full whole cell lysate in reducing sample buffer onto a 4–15% tris-glycine gel. Proteins were transferred to PVDF and then blocked in blocking buffer (TBST+5% non-fat milk) for one hour at room temperature. PKC alpha was detected at approximately 42 kDa using a PKA alpha Monoclonal Antibody (6D2.1) (Product # MA1-19732) at a dilution of 1:500 overnight at 4°C on a rocking platform, followed by a goat anti-mouse IgG Horseradish Peroxidase-conjugated Antibody at a dilution of 1:2,500 for one hour at room temperature. Chemiluminescent detection was performed.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to residues ESPAQNTAHLDQFERIK of human proteinkinase Ac alpha (PKAc alpha).|
|Storage buffer||PBS, pH 7.4|
|Contains||15mM sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Dot blot (DB)||Assay Dependent|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody reacts with human protein kinase A catalytic (PKAc) alpha subunit, and weakly with PKAc gamma subunit (both around 40 kDa). The recognized epitope of PKAc alpha is identical between man, sheep, pig, ox and dog.
MA1-19732 detects PKAc alpha in WB application with human samples.
Protein kinase A (PKA, cAMP-dependent protein kinase) is a key element of a ubiquitous signaling pathway important in the cell cycle, cellular communication, memory formation and behavior. PKA is composed of two catalytic (PKAc; Protein Kinase A catalytic subunit) and two regulatory subunits (PKAr). Upon binding cAMP, the complex dissociates to PKAr dimer and two activated PKAc ser/thr protein kinase catalytic monomers. The released PKAc can translocate into the nucleus and exert a regulatory role in the activation of multiple nuclear hormone receptors. However, PKAc-mediated activation of tonicity-dependent gene expression is cAMP independent. Humans express three types of PKAc subunit - PKAc alpha is present in most human tissues, PKAc beta and gamma are tissue-specific, the later is found in testes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.