Anti-PKR 抗体 [EPR19374] (ab184257)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19374] to PKR
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-PKR antibody [EPR19374]
PKR 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR19374] to PKR -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), ICC/IF, IP, WBmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: A459, K562, HeLa, Jurkat, 4T1, MCF7, HepG2 and bEnd.3 whole cell lysates. Mouse brain, cerebral cortex, hippocampus, lung, thymus and heart lysates; Rat brain, cerebral cortex, heart and spleen lysates; ICC/IF: bEnd.3 and A459 cells. Flow Cyt (Intra): bEnd.3 cells. IP: Mouse hippocampus lysate.
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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.
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 long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 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
EPR19374 -
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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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
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ab184257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) |
1/120.
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ICC/IF |
1/100.
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IP |
1/40.
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WB | (1) |
1/2000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
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特記事項 |
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Flow Cyt (Intra)
1/120. |
ICC/IF
1/100. |
IP
1/40. |
WB
1/2000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa). |
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
- Entrez Gene: 19106 Mouse
- Entrez Gene: 54287 Rat
- Omim: 176871 Human
- SwissProt: P19525 Human
- SwissProt: Q03963 Mouse
- SwissProt: Q63184 Rat
- Unigene: 131431 Human
see all -
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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All lanes : Anti-PKR antibody [EPR19374] (ab184257) at 1/1000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 5 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 58 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Lanes 1-5: Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.
ab184257 Anti-PKR antibody [EPR19374] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab184257 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-PKR antibody [EPR19374] (ab184257) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : EIF2AK2 knockout A549 cell lysate
Lane 3 : K-562 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 58 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.
ab184257 Anti-PKR antibody [EPR19374] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267000 (knockout cell lysate ab256901) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab184257 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: EIF2AK2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: K562 whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab184257 was shown to specifically react with EIF2AK2 when EIF2AK2 knockout samples were used. Wild-type and EIF2AK2 knockout samples were subjected to SDS-PAGE. Ab184257 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Anti-PKR antibody [EPR19374] (ab184257) at 1/1000 dilution + bEnd.3 (mouse brain endothelial cell) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 58 kDa
Exposure time: 114 secondsBlocking and diluting buffer: 5% NFDM/TBST
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized bEnd.3 (mouse brain endothelial cell) labeling PKR with ab184257 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preabsorbed (ab150081) secondary antibody at 1/1000 dilution (green).
Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 was used as a counterstain
The nuclear counterstain is DAPI (blue)
Confocal image showing cytoplasmic and weak nuclear staining in bEnd.3 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma epithelial cell) labeling PKR with ab184257 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preabsorbed (ab150081) secondary antibody at 1/1000 dilution (green).
Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 was used as a counterstain
The nuclear counterstain is DAPI (blue)
Confocal image showing cytoplasmic and weak nuclear staining in A549 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilised bEnd.3 (mouse brain endothelial cell) cells labeling PKR with ab184257 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.
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PKR was immunoprecipitated from mouse hippocampus lysate with ab184257 at 1/30 dilution (2μg in 0.35mg lysates).
Western blot was performed from the immunoprecipitate using ab184257 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: Mouse hippocampus tissue lysate 10 μg (Input).
Lane 2: ab184257 IP in Mouse hippocampus tissue lysate.
Lane 3: Rabbit IgG,monoclonal [EPR19374] - Isotype Control (ab172730) instead of ab184257 in mouse hippocampus tissue lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 31 seconds.
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All lanes : Anti-PKR antibody [EPR19374] (ab184257) at 1/2000 dilution
Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 58 kDa
Observed band size: 58 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-PKR antibody [EPR19374] (ab184257) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse cerebral cortex lysate
Lane 3 : Mouse hippocampus lysate
Lane 4 : Mouse lung lysate
Lane 5 : Mouse thymus lysate
Lane 6 : Rat brain lysate
Lane 7 : Rat cerebral cortex lysate
Lane 8 : 4T1 (Mouse mammary gland carcinoma cell line) whole cell lysate
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: 58 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,2,3 and 4: 3 minutes; Lane 5: 30 seconds; Lane 6,7 and 8: 10 seconds.
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All lanes : Anti-PKR antibody [EPR19374] (ab184257) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat heart lysate
Lane 5 : Rat spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 58 kDa
Observed band size: 58 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 3 minutes; Lane 3,4 and 5: 5 seconds.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (10)
ab184257 は 10 報の論文で使用されています。
- 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
- Tang J et al. Increased Mobile Zinc Regulates Retinal Ganglion Cell Survival via Activating Mitochondrial OMA1 and Integrated Stress Response. Antioxidants (Basel) 11:N/A (2022). PubMed: 36290724
- Dou S et al. Ubiquitination and degradation of NF90 by Tim-3 inhibits antiviral innate immunity. Elife 10:N/A (2021). PubMed: 34110282
- Yang SR et al. LCC18, a benzamide-linked small molecule, ameliorates IgA nephropathy in mice. J Pathol 253:427-441 (2021). PubMed: 33373038
- Gabaev I et al. Quantitative Proteomics Analysis of Lytic KSHV Infection in Human Endothelial Cells Reveals Targets of Viral Immune Modulation. Cell Rep 33:108249 (2020). PubMed: 33053346
- Wu CH et al. A Synthetic Small Molecule F240B Decreases NLRP3 Inflammasome Activation by Autophagy Induction. Front Immunol 11:607564 (2020). PubMed: 33424855
- Lee H et al. Cell Type-Specific Transcriptomics Reveals that Mutant Huntingtin Leads to Mitochondrial RNA Release and Neuronal Innate Immune Activation. Neuron 107:891-908.e8 (2020). PubMed: 32681824
- Singh VP et al. Persistent DNA Damage and Senescence in the Placenta Impacts Developmental Outcomes of Embryos. Dev Cell 54:333-347.e7 (2020). PubMed: 32800293
- Cao Y et al. ER stress-induced mediator C/EBP homologous protein thwarts effector T cell activity in tumors through T-bet repression. Nat Commun 10:1280 (2019). PubMed: 30894532
- Chiu HW et al. Glucosamine inhibits IL-1ß expression by preserving mitochondrial integrity and disrupting assembly of the NLRP3 inflammasome. Sci Rep 9:5603 (2019). PubMed: 30944389