Recombinant 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: WB, IP
- Reacts with: Human, Pig
Related conjugates and formulations
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 -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC-P -
Species reactivity
Reacts with: Human, Pig -
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. IP: HeLa
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General notes
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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...
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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.
Application | Abreviews | Notes |
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WB | (6) |
1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
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IP |
1/10.
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Notes |
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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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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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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? -
PKR was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A(100nM 30min) whole cell lysate 10 µg with ab32036 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A(100nM 30min) whole cell lysate 10 µg
Lane 2: ab32036 IP in HeLa treated with Calyculin A(100nM 30min) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab32036 in HeLa treated with Calyculin A(100nM 30min) whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Lower bands could be cleavage form. (PMID:28702377)
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.
Datasheets and documents
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SDS download
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Datasheet download
References (107)
ab32036 has been referenced in 107 publications.
- Price AM et al. Adenovirus prevents dsRNA formation by promoting efficient splicing of viral RNA. Nucleic Acids Res 50:1201-1220 (2022). PubMed: 34671803
- Breuer J et al. What goes around comes around: Artificial circular RNAs bypass cellular antiviral responses. Mol Ther Nucleic Acids 28:623-635 (2022). PubMed: 35497503
- Lin WY et al. Repositioning of the Angiotensin II Receptor Antagonist Candesartan as an Anti-Inflammatory Agent With NLRP3 Inflammasome Inhibitory Activity. Front Immunol 13:870627 (2022). PubMed: 35669789
- Eo H & Valentine RJ Saturated Fatty Acid-Induced Endoplasmic Reticulum Stress and Insulin Resistance Are Prevented by Imoxin in C2C12 Myotubes. Front Physiol 13:842819 (2022). PubMed: 35936891
- Chludzinski E et al. Phenotypic and Transcriptional Changes of Pulmonary Immune Responses in Dogs Following Canine Distemper Virus Infection. Int J Mol Sci 23:N/A (2022). PubMed: 36077417