Protein Kinase A regulatory subunit I alpha Knockout 293T Cell Lysate Summary
Preparation Method |
Knockout achieved by using CRISPR/Cas9,22 bp deletion in exon 1 and 5 bp deletion in exon 1 and 1 bp insertion in exon 1 |
| Gene |
PRKAR1A |
Applications/Dilutions
| Dilutions |
|
| Application Notes |
You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles. |
Packaging, Storage & Formulations
| Storage |
Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles. |
| Buffer |
0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines. |
| Concentration |
LYOPH |
| Reconstitution Instructions |
To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel. |
Lysate Details for Array
| Type |
Knockout 293T Cell |
| Life Stage |
Embryonic |
| Tissue |
Kidney |
Notes
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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.
Alternate Names for Protein Kinase A regulatory subunit I alpha Knockout 293T Cell Lysate
Background
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. This gene encodes one of the regulatory subunits. This protein was found to be a tissue-specific extinguisher that down-regulates the expression of seven liver genes in hepatoma x fibroblast hybrids. Mutations in this gene cause Carney complex (CNC). This gene can fuse to the RET protooncogene by gene rearrangement and form the thyroid tumor-specific chimeric oncogene known as PTC2. A nonconventional nuclear localization sequence (NLS) has been found for this protein which suggests a role in DNA replication via the protein serving as a nuclear transport protein for the second subunit of the Replication Factor C (RFC40). Several alternatively spliced transcript variants encoding two different isoforms have been observed. [provided by RefSeq, Jan 2013]
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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