Anti-PKR 抗体 [Y117] (ab32506)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y117] to PKR
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-PKR antibody [Y117]
PKR 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [Y117] to PKR -
由来種
Rabbit -
特異性
This antibody does not cross-react with other GCN2 family members.
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アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide within Human PKR aa 500-600 (C terminal). The exact sequence is proprietary.
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ポジティブ・コントロール
- WB: MCF-7, HEK293 and HeLa whole cell lysate (ab150035) and human liver carcinoma tissue lysate. ICC/IF: Wild type HAP1, MCF-7 and HeLa cells. IHC-P: Human liver carcinoma and colon carcinoma tissue.
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特記事項
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
Y117 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Isotype control
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab32506の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB | (3) |
1/5000 - 1/20000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
For unpurified use at 1/250 - 1/500 |
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IP |
1/80 - 1/100.
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特記事項 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/5000 - 1/20000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. For unpurified use at 1/250 - 1/500 |
IP
1/80 - 1/100. |
ターゲット情報
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機能
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. -
配列類似性
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. -
翻訳後修飾
Autophosphorylated 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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参照データベース
- Entrez Gene: 5610 Human
- Omim: 176871 Human
- SwissProt: P19525 Human
- Unigene: 131431 Human
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別名
- Double stranded RNA activated protein kinase; antibody
- E2AK2_HUMAN antibody
- eIF-2A protein kinase 2 antibody
see all
画像
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Lanes 1, 3 and 5: PKR knockout HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: Wild-type HAP1 cell lysate (20 µg)
Lanes 1 and 2: Green signal from target - ab32506 observed at 62 kDa
Lanes 3 and 4: Red signal from loading control - ab8245 observed at 37 kDa
Lanes 5 and 6: Merged (red and green) signal
ab32506 was shown to specifically react with PKR when PKR knockout samples were used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32506 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 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 1/10 000 dilution for 1 h at room temperature before imaging. -
ab32506 staining PKR in wild-type HAP1 cells (top panel) and PKR knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32506 at 1/400 dilution and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.
This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 4% formaldehyde (10 min).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -
All lanes : Anti-PKR antibody [Y117] (ab32506) at 1/20000 dilution
Lane 1 : MCF-7 (human breast carcinoma) whole cell lysates
Lane 2 : HEK293 (human embryonic kidney) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 62 kDa
Additional bands at: 68 kDa. We are unsure as to the identity of these extra bands.Purified format.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] (ab32506)
ab32506 staining PKR in human liver carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control 1: PBS in place of primary antibody.
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Intracellular Flow Cytometry analysis of MCF-7 (human breast carcinoma) cells labeling PKR with purified ab32506 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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Anti-PKR antibody [Y117] (ab32506) at 1/10000 dilution (unpurified) + MCF-7 cell lysate
Predicted band size: 62 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted? -
ab32506 immunoprecipitating PKR. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.
Lane 1: HEK293 (human embryonic kidney) whole cell lysate (10ug)
Lane 2: HEK293 (human embryonic kidney) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab32506 in HEK293 (human embryonic kidney) whole cell lysate -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] (ab32506)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using unpurified ab32506 at 1/100 dilution.
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ab32506 staining PKR in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab32506 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.
Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) -
Immunofluorescent staining of HeLa cells using unpurified ab32506 at 1/250 dilution.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (29)
ab32506 は 29 報の論文で使用されています。
- Zhu Y et al. Polynucleotide phosphorylase protects against renal tubular injury via blocking mt-dsRNA-PKR-eIF2α axis. Nat Commun 14:1223 (2023). PubMed: 36869030
- Khilazheva ED et al. Impact of NLRP3 Depletion on Aging-Related Metaflammation, Cognitive Function, and Social Behavior in Mice. Int J Mol Sci 24:N/A (2023). PubMed: 38068904
- Price AM et al. Adenovirus prevents dsRNA formation by promoting efficient splicing of viral RNA. Nucleic Acids Res 50:1201-1220 (2022). PubMed: 34671803
- Zhao J et al. Role of Stress Granules in Suppressing Viral Replication by the Infectious Bronchitis Virus Endoribonuclease. J Virol 96:e0068622 (2022). PubMed: 35638780
- McFadden MJ et al. FTO Suppresses STAT3 Activation and Modulates Proinflammatory Interferon-Stimulated Gene Expression. J Mol Biol 434:167247 (2022). PubMed: 34537236