Product nameAnti-PACT (PKR activating protein) / PRKRA antibody
See all PACT (PKR activating protein) / PRKRA primary antibodies
DescriptionRabbit polyclonal to PACT (PKR activating protein) / PRKRA
Tested applicationsSuitable for: ICC/IF, WB, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Cow
- HeLa Whole Cell Lysate Testis (Mouse) Tissue Lysate PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab31967 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).|
|IP||Use a concentration of 5 µg/ml.|
FunctionActivates 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 diseaseDefects 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 similaritiesBelongs to the PRKRA family.
Contains 3 DRBM (double-stranded RNA-binding) domains.
DomainSelf-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.
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 localizationCytoplasm > perinuclear region.
- Information by UniProt
- DYT16 antibody
- HSD14 antibody
- Interferon inducible double stranded RNA dependent protein kinase activator A antibody
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PACT (PKR activating protein)/PRKRA knockout HAP1 cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab31967 observed at 36 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab31967 was shown to recognize PACT (PKR activating protein)/PRKRA when PACT (PKR activating protein)/PRKRA knockout samples were used, along with additional cross-reactive bands. Wild-type and PACT (PKR activating protein)/PRKRA knockout samples were subjected to SDS-PAGE. ab31967 and ab18058 (loading control to Vinculin) were diluted at 1 μg/ml and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.
ICC/IF image of ab31967 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31967, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967) at 1 µg/ml
Lane 1 : Testis (Mouse) Tissue Lysate - normal tissue
Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.
PACT (PKR activating protein) / PRKRA was immunoprecipitated using 0.5mg Mouse Testis tissue, 5µg of Rabbit polyclonal to PACT (PKR activating protein) / PRKRA and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Testis tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31967.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 34kDa; PACT (PKR activating protein) / PRKRA
This product has been referenced in:
- Warner MJ et al. S6K2-mediated regulation of TRBP as a determinant of miRNA expression in human primary lymphatic endothelial cells. Nucleic Acids Res 44:9942-9955 (2016). WB ; Human . Read more (PubMed: 27407113) »
- Wheatley AK et al. Co-Expression of miRNA Targeting the Expression of PERK, but Not PKR, Enhances Cellular Immunity from an HIV-1 Env DNA Vaccine. PLoS One 6:e18225 (2011). WB ; Human . Read more (PubMed: 21464971) »