Human PRKRA (PACT) knockout HEK293T cell pellet (ab279114)
Overview
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Product name
Human PRKRA (PACT) knockout HEK293T cell pellet
See all PACT (PKR activating protein) / PRKRA kits -
Product overview
Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.
Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 2 bp deletion in exon1. -
Passage number
Knockout validation
Sanger Sequencing, Western Blot (WB)Notes
Pellet size: 5 million cells/vial.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit Human PRKRA knockout HEK293T cell pellet 1 vial Human wild-type HEK293T cell pellet 1 vial -
Research areas
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Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Target
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Function
Activates EIF2AK2/PKR in the absence of double stranded RNA (dsRNA), leading to phosphorylation of EIF2S1/EFI2-alpha and inhibition of translation and induction of apoptosis. Required for siRNA production by DICER1 and for subsequent siRNA-mediated post-transcriptional gene silencing. Does not seem to be required for processing of pre-miRNA to miRNA by DICER1. -
Involvement in disease
Defects in PRKRA are the cause of dystonia type 16 (DYT16) [MIM:612067]. DYT16 is an early-onset dystonia-parkinsonism disorder. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYT16 patients have progressive, generalized dystonia with axial muscle involvement, oro-mandibular (sardonic smile) and laryngeal dystonia and, in some cases, parkinsonian features. -
Sequence similarities
Belongs to the PRKRA family.
Contains 3 DRBM (double-stranded RNA-binding) domains. -
Domain
Self-association may occur via interactions between DRBM domains as follows: DRBM 1/DRBM 1, DRBM 1/DRBM 2, DRBM 2/DRBM 2 or DRBM 3/DRBM3. -
Post-translational
modificationsPhosphorylated at Ser-246 in unstressed cells and at Ser-287 in stressed cells. Phosphorylation at Ser-246 appears to be a prerequisite for subsequent phosphorylation at Ser-287. Phosphorylation at Ser-246 and Ser-287 are necessary for activation of EIF2AK2/PKR under conditions of stress. -
Cellular localization
Cytoplasm > perinuclear region. - Information by UniProt
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Alternative names
- DYT16
- HSD14
- Interferon inducible double stranded RNA dependent protein kinase activator A
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit Human PRKRA knockout HEK293T cell pellet 1 vial Human wild-type HEK293T cell pellet 1 vial -
Research areas
-
Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Images
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Western blot - Human PRKRA (PACT) knockout HEK293T cell pellet (ab279114)
Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:PRKRA knockout HeLa cell lysate (20 ug)
Lane 3:K-562 cell lysate (20 ug)
Lane 4:HepG2 cell lysate (20 ug)ab31967 was shown to specifically react with PACT in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266806 (knockout cell lysate ab258141) was used. Wild-type and PACT knockout samples were subjected to SDS-PAGE. ab31967 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Human PRKRA (PACT) knockout HEK293T cell pellet (ab279114)Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:PRKRA knockout HeLa cell lysate (20 ug)
Lane 3:K-562 cell lysate (20 ug)
Lane 4:HepG2 cell lysate (20 ug)ab75749 was shown to specifically react with PACT in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266806 (knockout cell lysate ab258141) was used. Wild-type and PACT knockout samples were subjected to SDS-PAGE. ab75749 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human PRKRA (PACT) knockout HEK293T cell pellet (ab279114)Allele-1: 2 bp deletion in exon1
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Sanger Sequencing - Human PRKRA (PACT) knockout HEK293T cell pellet (ab279114)Allele-2: 1 bp deletion in exon1