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Product Info Summary
| SKU: | M00653-2 |
|---|---|
| Size: | 100 μl/vial |
| Reactive Species: | Human, Monkey, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IP, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-PRKACA Antibody (Monoclonal, 33P21)
SKU/Catalog Number
M00653-2
Size
100 μl/vial
Form
Liquid
Description
Boster Bio Anti-PRKACA Antibody (Monoclonal, 33P21) catalog # M00653-2. Tested in WB, IHC, ICC/IF, IP, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat, Monkey.
Storage & Handling
Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-PRKACA Antibody (Monoclonal, 33P21) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00653-2)
Host
Rabbit
Contents
Rabbit IgG in stabilizing components, phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Monoclonal
Clone Number
33P21
Immunogen
Recombinant protein within human PKA C-alpha aa 3-349.
Reactive Species
M00653-2 is reactive to PRKACA in Human, Monkey, Mouse, Rat
Observed Molecular Weight
38 kDa
Calculated molecular weight
40.6 kDa
Background of PRKACA
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.
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
M00653-2 is guaranteed for Flow Cytometry, IP, IF, IHC, 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, 1:500-2000
Immunohistochemistry, 1:50-200
Immunocytochemistry/Immunofluorescence, 1:50-200
ImmunoPrecipitation, 1:50
Flow Cytometry (Fixed), 1:50-200
Positive Control
WB: human PC-3 whole cell, human MCF-7 whole cell, human Hela whole cell, human U251 whole cell, rat heart tissue, rat brain tissue, mouse heart tissue, mouse brain tissue
IHC: human testis cancer tissue
Validation Images & Assay Conditions
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Western blot analysis of PRKACA using anti-PRKACA antibody (M00653-2).
Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human PC-3 whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat heart tissue lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse heart tissue lysates,
Lane 8: mouse brain 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-PRKACA antigen affinity purified monoclonal antibody (M00653-2) at 1:1000 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for PRKACA at approximately 38 kDa. The expected band size for PRKACA is at 41 kDa.
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IHC analysis of PRKACA using anti-PRKACA antibody (M00653-2).
PRKACA was detected in a paraffin-embedded section of human testis cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:200 rabbit anti-PRKACA Antibody (M00653-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Specific Publications For Anti-PRKACA Antibody (Monoclonal, 33P21) (M00653-2)
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