Human PRKRA (PACT) knockout HEK-293T cell lysate
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PRKRA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 2 bp deletion in exon1.
View Alternative Names
DYT16, HSD14, Interferon-inducible double stranded RNA-dependent protein kinase activator A, PACT, PKR-associated protein X, PKR-associating protein X, PRKRA_HUMAN, Protein activator of the interferon-induced protein kinase, Protein kinase, Protein kinase interferon inducible double stranded RNA dependent activator, RAX, interferon-inducible double stranded RNA-dependent activator
- WB
Unknown
Western blot - Human PRKRA (PACT) knockout HEK-293T cell lysate (AB258141)
Lane 1 : Wild-type HEK-293T cell lysate (20 ug)
Lane 2 : PRKRA knockout HEK-293T 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 HEK-293T 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.
All lanes:
Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody (<a href='/en-us/products/primary-antibodies/pact-pkr-activating-protein-prkra-antibody-ab31967'>ab31967</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PRKRA knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-prkra-pact-knockout-hek-293t-cell-line-ab266806'>ab266806</a>)
Lane 3:
K-562 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 34 kDa
Observed band size: 36 kDa
false
- WB
Lab
Western blot - Human PRKRA (PACT) knockout HEK-293T cell lysate (AB258141)
Lane 1 : Wild-type HEK-293T cell lysate (20 ug)
Lane 2 : PRKRA knockout HEK-293T 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 HEK-293T 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.
All lanes:
Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (<a href='/en-us/products/primary-antibodies/pact-pkr-activating-protein-prkra-antibody-epr3224-ab75749'>ab75749</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PRKRA knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-prkra-pact-knockout-hek-293t-cell-line-ab266806'>ab266806</a>)
Lane 3:
K-562 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 34 kDa
Observed band size: 36 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PRKRA (PACT) knockout HEK-293T cell lysate (AB258141)
Allele-2 : 1 bp deletion in exon1
- Sanger seq
Unknown
Sanger Sequencing - Human PRKRA (PACT) knockout HEK-293T cell lysate (AB258141)
Allele-1 : 2 bp deletion in exon1
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
PACT involves in the cellular stress response signaling. It serves as an activator for PKR a kinase that becomes active during viral infections and stress. PACT forms complexes with PKR facilitating its activation and subsequent initiation of downstream signaling cascades. It contributes significantly to the regulation of protein synthesis and cell growth by modulating these pathways. In conditions of stress PACT ensures adequate PKR activation for host defense.
Pathways
PACT is key in the interferon signaling pathway and the stress response pathway. It partners with PKR to suppress protein synthesis through phosphorylation of the eukaryotic initiation factor 2 alpha (eIF2α) which slows down protein production during cellular stress. This interaction places PACT at a junction of antiviral defense and stress adaptation with PKR serving as an essential protein involved in these pathways.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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