使用敲除细胞株进行验证

Anti-PACT (PKR activating protein) / PRKRA抗体(ab31967)

概述

  • 产品名称Anti-PACT (PKR activating protein) / PRKRA抗体
    参阅全部 PACT (PKR activating protein) / PRKRA 一抗
  • 描述
    兔多克隆抗体to PACT (PKR activating protein) / PRKRA
  • 经测试应用适用于: ICC/IF, WB, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Cow
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human PACT (PKR activating protein)/ PRKRA.

    (Peptide available as ab30768.)

  • 阳性对照
    • HeLa Whole Cell Lysate Testis (Mouse) Tissue Lysate PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

性能

相关产品

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能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.
  • 疾病相关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.
  • 序列相似性Belongs to the PRKRA family.
    Contains 3 DRBM (double-stranded RNA-binding) domains.
  • 结构域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.
  • 翻译后修饰Phosphorylated 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.
  • 细胞定位Cytoplasm > perinuclear region.
  • Information by UniProt
  • 数据库链接
  • 别名
    • DYT16 antibody
    • HSD14 antibody
    • Interferon inducible double stranded RNA dependent protein kinase activator A antibody
    • interferon-inducible double stranded RNA-dependent activator antibody
    • Interferon-inducible double stranded RNA-dependent protein kinase activator A antibody
    • PACT antibody
    • PKR associated protein X antibody
    • PKR associating protein X antibody
    • PKR-associated protein X antibody
    • PKR-associating protein X antibody
    • PRKRA antibody
    • PRKRA_HUMAN antibody
    • Protein activator of the interferon induced protein kinase antibody
    • Protein activator of the interferon-induced protein kinase antibody
    • Protein kinase antibody
    • Protein kinase interferon inducible double stranded RNA dependent activator antibody
    • RAX antibody
    see all

Anti-PACT (PKR activating protein) / PRKRA antibody 图像



  • Predicted band size : 34 kDa

    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 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 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).



  • Performed under reducing conditions.

    Predicted band size : 34 kDa
  • 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.

    Secondary
    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

Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967)参考文献

This product has been referenced in:
  • 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) »

See 1 Publication for this product

Product Wall

There are currently no Abreviews or Questions for ab31967.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"