The catalytic subunit C-alpha of PKA (PKA C alpha ) is a member of the Ser/Thr protein kinase family and has been assigned to chromosome region 19p13.1 (1). Null mutation in PKA C alpha leads to early postnatal lethality in the majority of C-alpha knockout mice. Surprisingly, a small percentage of C-alpha knockout mice, although runted, survived to adulthood. In these animals, compensatory increases in C-beta levels occurred in brain whereas many tissues, including skeletal muscle, heart, and sperm contained less than 10% of the normal PKA activity (2).
Recombinant Human Active PKA C alpha Protein, CF
R&D Systems | Catalog # 4268-KS
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Key Product Details
- R&D Systems Sf 9 (baculovirus)-derived Recombinant Human Active PKA C alpha Protein (4268-KS)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
Sf 9 (baculovirus)
Accession Number
Applications
Bioactivity
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Product Specifications
Source
Spodoptera frugiperda, Sf 9 (baculovirus)-derived human PKA C alpha protein
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
N-terminal Sequence Analysis
Using an N-terminal GST tag
SDS-PAGE
69 kDa
Activity
The activity of PKA C alpha is typically 1626-2201 nmol/min/mg using a synthetic peptide substrate.
Reviewed Applications
Read 3 reviews rated 4.7 using 4268-KS in the following applications:
Scientific Data Images for Recombinant Human Active PKA C alpha Protein, CF
Recombinant Human Active PKA C alpha Protein SDS-PAGE.
The approximate molecular weight is 69 kDa and the purity is > 90%.Formulation, Preparation, and Storage
4268-KS
| Formulation | Supplied in 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 10 mM Glutathione, 0.25 mM DTT, 0.1 mM EDTA, 0.1 mM PMSF, and 25% Glycerol. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | This product is stable at ≤ ‑70 °C for up to 1 year from the date of receipt. For optimal storage, aliquot into smaller quantities after centrifugation and store at recommended temperature. Avoid repeated freeze-thaw cycles. |
Background: PKA C alpha
References
- Tasken, K. et al. (1996) Genomics 36:535.
- Skalhegg, B.S. et al. (2002) Molec. Endocr. 16:630.
Long Name
cAMP-dependent Protein Kinase Catalytic Subunit alpha
Alternate Names
Cs, Pkaca, PKCA1, PKCD, PRKACA
Gene Symbol
PRKACA
UniProt
Additional PKA C alpha Products
Product Documents for Recombinant Human Active PKA C alpha Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Active PKA C alpha Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Active PKA C alpha Protein, CF
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4.7 out of 5
3 Customer Ratings
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Application: Cell ProliferationVerified Customer | Posted 09/04/2019
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Protocols
View specific protocols for Recombinant Human Active PKA C alpha Protein, CF (4268-KS):
Materials
- Active Kinase - Active PKA C alpha (0.1 μg/μL) diluted with Kinase Dilution Buffer VII to the concentrations indicated in Figure 2. Note: These are suggested working dilutions. Optimal dilutions should be determined by each laboratory for each application.
- Kinase Assay Buffer I - 25 mM MOPS, pH 7.2, 12.5 mM beta -glycerolphosphate, 25 mM MgCl2, 5 mM EGTA, 2 mM EDTA. Add 0.25 mM DTT to the Kinase Assay Buffer prior to use.
- Kinase Dilution Buffer VII - Kinase Assay Buffer I diluted at a 1:4 ration (5X dilution) with 50 ng/μL BSA and 5% glycerol solution.
- 10 mM ATP Stock Solution - Prepare the ATP Stock Solution by dissolving 55 mg of ATP in 10 mL of Kinase Assay Buffer I. Store 200 μL aliquots at ≤ -20° C.
- [33P]-ATP Assay Cocktail - Prepare 250 μM [33P]-ATP Assay Cocktail in a designated radioactive work area by combining 150 μL of 10 mM ATP Stock Solution, 100 μL of [33P]-ATP (1 mCi/100 μL), and 5.75 mL of Kinase Assay Buffer. Store 1 mL aliquots at ≤ -20 °C.
- Substrate - CREBtide synthetic peptide substrate (KRREILSRRPSYR) diluted in distilled water to a final concentration of 1 mg/mL.
- Thaw the [33P]-ATP Assay Cocktail in a shielded container in a designated radioactive work area.
- Thaw the Active PKA C alpha, Kinase Assay Buffer, Substrate, and Kinase Dilution Buffer on ice.
- In a pre-cooled microfuge tube, add the following reaction components bringing the initial reaction volume up to 20 μL.
a. Diluted Active PKA C alpha : 10 μL
b. Stock solution of Substrate (1 mg/mL): 5 μL
c. Distilled water (2-8 °C): 5 μL - Set up the blank control as outlined in Step 3, excluding the addition of the substrate. Replace the substrate with an equal volume of distilled or deionized water.
- Initiate the reaction with the addition of 5 μL [33P]-ATP Assay Cocktail, bringing the final volume up to 25 μL. Incubate the mixture in a water bath at 30 °C for 15 minutes.
- After the 15 minute incubation, terminate the reaction by spotting 20 μL of the reaction mixture onto individual pre-cut strips of phosphocellulose P81 paper.
- Air dry the pre-cut P81 strip and sequentially wash in a 1% phosphoric acid solution (add 10 mL of phosphoric acid to 990 mL of distilled water) with constant gentle stirring. It is recommended that the strips be washed a total of three times for approximately 10 minutes each.
- Count the radioactivity on the P81 paper in the presence of scintillation fluid in a scintillation counter.
- Determine the corrected cpm by subtracting the blank control value (see Step 4) for each sample and calculate the kinase specific activity as outlined below.
Calculation of [33P]-ATP Specific Activity (SA) (cpm/pmol)
Specific Activity (SA) = cpm for 5 μL [33P]-ATP/pmol of ATP (in 5 μL of a 250 μM ATP stock solution; i.e. 1250 pmol)
Calculation of Kinase Specific Activity (SA) (pmol/minutes/μg or nmol/minutes/mg)
Corrected cpm from reaction / [(SA of 33P-ATP in cpm/pmol) x (Reaction time in minutes) x (Enzyme amount in μg or mg)] x [(Reaction volume) / (Spot Volume)]
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