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Product Info Summary
| SKU: | A03911-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-PKA R2/PKR2/PRKAR2A Antibody Picoband®
SKU/Catalog Number
A03911-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-PKA R2/PKR2/PRKAR2A Antibody Picoband® catalog # A03911-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-PKA R2/PKR2/PRKAR2A Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03911-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human PKA R2/PKR2/PRKAR2A recombinant protein (Position: E20-K315).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03911-2 is reactive to PRKAR2A in Human, Mouse, Rat
Observed Molecular Weight
52 kDa
Calculated molecular weight
45.5 kDa
Background of PRKAR2A
cAMP-dependent protein kinase type II-alpha regulatory subunit is an enzyme that in humans is encoded by the PRKAR2A gene. cAMP is a signaling molecule important for a variety of cellular functions. cAMP exerts its effects by activating the cAMP-dependent protein kinase, which transduces the signal through phosphorylation of different target proteins. The inactive kinase holoenzyme is a tetramer composed of two regulatory and two catalytic subunits. 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. The protein encoded by this gene is one of the regulatory subunits. This subunit can be phosphorylated by the activated catalytic subunit. It may interact with various A-kinase anchoring proteins and determine the subcellular localization of cAMP-dependent protein kinase. This subunit has been shown to regulate protein transport from endosomes to the Golgi apparatus and further to the endoplasmic reticulum (ER).
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A03911-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.25 μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human K562 whole cell, human Jurkat whole cell, human Hela whole cell, human U20S whole cell, rat brain tissue, mosue brain tissue, mouse lung tissue
ICC/IF: Hela cell
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of PKA R2/PKR2/PRKAR2A using anti-PKA R2/PKR2/PRKAR2A antibody (A03911-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human U20S whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: mosue brain tissue lysates,
Lane 7: mouse lung tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PKA R2/PKR2/PRKAR2A antigen affinity purified polyclonal antibody (Catalog # A03911-2) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PKA R2/PKR2/PRKAR2A at approximately 52 kDa. The expected band size for PKA R2/PKR2/PRKAR2A is at 52 kDa.
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IF analysis of PKA R2/PKR2/PRKAR2A using anti-PKA R2/PKR2/PRKAR2A antibody (A03911-2).
PKA R2/PKR2/PRKAR2A was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-PKA R2/PKR2/PRKAR2A Antibody (A03911-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of A431 cells using anti-PKA R2/PKR2/PRKAR2A antibody (A03911-2).
Overlay histogram showing A431 cells stained with A03911-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PKA R2/PKR2/PRKAR2A Antibody (A03911-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-PKA R2/PKR2/PRKAR2A Antibody Picoband® (A03911-2)
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