Anti-PKA alpha + beta (catalytic subunits) (phospho T197) antibody (ab5815)
Key features and details
- Rabbit polyclonal to PKA alpha + beta (catalytic subunits) (phospho T197)
- Suitable for: WB
- Reacts with: Mouse, Recombinant fragment
- Isotype: IgG
Overview
-
Product name
Anti-PKA alpha + beta (catalytic subunits) (phospho T197) antibody
See all PKA alpha + beta (catalytic subunits) primary antibodies -
Description
Rabbit polyclonal to PKA alpha + beta (catalytic subunits) (phospho T197) -
Host species
Rabbit -
Specificity
This antibody exibited a preference for PKA catalytic subunit beta in some tested cell lines. -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Mouse, Recombinant fragment
Predicted to work with: Cow, Pig
-
Immunogen
Synthetic phosphopeptide derived from a region of human PKA that contains threonine 197.
-
Positive control
- Forskolin-treated NIH3T3 cells, and Y-1 mouse adrenal cortical cells.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated PKA. The final product is generated by affinity chromatography using a PKA-derived peptide that is phosphorylated at threonine 197. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
-
Western blot - Anti-PKA alpha + beta (catalytic subunits) (phospho T197) antibody (ab5815)Peptide Competition and Phosphatase Treatment: Lysates prepared from Y1 Adrenocortical cells were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% BSA-TBST buffer for two hours at room temperature, then incubated with 0.35
µ g/mL ab5815 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’ 2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignalTM method. The data show that the peptide corresponding to PKA [pT197] blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
Peptide Competition and Phosphatase Treatment: Lysates prepared from Y1 Adrenocortical cells were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% BSA-TBST buffer for two hours at room temperature, then incubated with 0.35 µg/mL ab5815 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’ 2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignalTM method. The data show that the peptide corresponding to PKA [pT197] blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
