Recombinant Anti-PKR (phospho T446) antibody [E120] (ab32036)
![Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)](https://www.abcam.com/ps/products/32/ab32036/Images/ab32036-395134-anti-pkr-phospho-t446-antibody-e120-western-blot-calyculin-a-tnf-alpha-treated-hela.jpg "Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E120] to PKR (phospho T446)
- Suitable for: IHC-P, WB, IP
- Reacts with: Human, Pig
Overview
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Product name
Anti-PKR (phospho T446) antibody [E120]
See all PKR primary antibodies -
Description
Rabbit monoclonal [E120] to PKR (phospho T446) -
Host species
Rabbit -
Specificity
The antibody only detects PKR phosphorylated on Threonine 446. -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P PigWB Human
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Immunogen
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Positive control
- WB: HeLa treated with Calyculin A and TNF-alpha whole cell lysate; Calynculin A and IFN treated HeLa cell lysate. IHC-P: Human colon tissue.
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General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E120 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32036 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| IHC-P |
Pig
|
| WB |
Human
|
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P |
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
| WB | (6) |
1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
|
| IP |
1/10.
|
| Notes |
|---|
|
IHC-P
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
|
WB
1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa). |
|
IP
1/10. |
Target
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Function
Following activation by double-stranded RNA in the presence of ATP, the kinase becomes autophosphorylated and can catalyze the phosphorylation of the translation initiation factor EIF2S1, which leads to an inhibition of the initiation of protein synthesis. Double-stranded RNA is generated during the course of a viral infection. -
Sequence similarities
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.
Contains 2 DRBM (double-stranded RNA-binding) domains.
Contains 1 protein kinase domain. -
Post-translational
modificationsAutophosphorylated on several Ser and Thr residues. Autophosphorylation of Thr-451 is dependent on Thr-446 and is stimulated by dsRNA binding and dimerization. Autophosphorylation apparently leads to the activation of the kinase. - Information by UniProt
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Database links
- Entrez Gene: 5610 Human
- Omim: 176871 Human
- SwissProt: P19525 Human
- Unigene: 131431 Human
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Alternative names
- Double stranded RNA activated protein kinase; antibody
- E2AK2_HUMAN antibody
- eIF-2A protein kinase 2 antibody
see all
Images
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Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)All lanes : Anti-PKR (phospho T446) antibody [E120] (ab32036) at 1/2000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma) treated with 100 ng/mL Calyculin A whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma) treated with 100 ng/mL Calyculin A followed by 20 ng/mL TNF alpha whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 62 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking/Diluting buffer: 5% NFDM/TBST
Positive control: Rabbit monoclonal [Y117] to PKR (ab32506).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR (phospho T446) antibody [E120] (ab32036)](https://www.abcam.com/ps/products/32/ab32036/Images/ab32036-43698-anti-pkr-phospho-t446-antibody-e120-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR (phospho T446) antibody [E120] (ab32036)Immunohistochemical analysis of PKR (phospho T446) expression in normal colon tissue, using 1/50 unpurified ab32036. Heat mediated antigen retrieval performed before IHC staining.
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![Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)](https://www.abcam.com/ps/products/32/ab32036/Images/ab32036-43699-anti-pkr-phospho-t446-antibody-e120-western-blot.jpg)
Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)All lanes : Anti-PKR (phospho T446) antibody [E120] (ab32036) at 1/1000 dilution (unpurified)
Lane 1 : Hela (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : Hela cell lysate + IFN + CalyculinA
Lane 3 : Hela cell lysate + Pptase
Predicted band size: 62 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted? -
![Anti-PKR (phospho T446) antibody [E120] (ab32036)](https://www.abcam.com/ps/products/32/ab32036/Images/ab32036-11-benefits-of-recombinant-antibodies.png)
Anti-PKR (phospho T446) antibody [E120] (ab32036)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
References (73)
ab32036 has been referenced in 73 publications.
- Cheng J et al. Typical Stress Granule Proteins Interact with the 3' Untranslated Region of Enterovirus D68 To Inhibit Viral Replication. J Virol 94:N/A (2020). PubMed: 31941779
- Wuerth JD et al. eIF2B as a Target for Viral Evasion of PKR-Mediated Translation Inhibition. mBio 11:N/A (2020). PubMed: 32665273
- Visser LJ et al. Foot-and-Mouth Disease Virus Leader Protease Cleaves G3BP1 and G3BP2 and Inhibits Stress Granule Formation. J Virol 93:N/A (2019). PubMed: 30404792
- Sauer M et al. DHX36 prevents the accumulation of translationally inactive mRNAs with G4-structures in untranslated regions. Nat Commun 10:2421 (2019). PubMed: 31160600
- Hood IV et al. Crystal structure of an adenovirus virus-associated RNA. Nat Commun 10:2871 (2019). PubMed: 31253805
