Anti-PACT (PKR activating protein) / PRKRA 抗体 (ab31967)
Key features and details
- Rabbit polyclonal to PACT (PKR activating protein) / PRKRA
- Suitable for: ICC/IF, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-PACT (PKR activating protein) / PRKRA antibody
PACT (PKR activating protein) / PRKRA 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to PACT (PKR activating protein) / PRKRA -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, WB, IPmore details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Cow
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免疫原
Synthetic peptide corresponding to Human PACT (PKR activating protein)/ PRKRA aa 100-200 conjugated to keyhole limpet haemocyanin.
(Peptide available asab30768) -
ポジティブ･コントロール
- WB: HEK-293T, K562, HepG2, HAP1 and PC12 whole cell lysates; Mouse testis tissue lysate. ICC/IF: HeLa cells. IP: Mouse testis tissue.
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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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
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ab31967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 |
|---|---|---|
| ICC/IF |
Use a concentration of 5 µg/ml.
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| WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
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| IP |
Use a concentration of 5 µg/ml.
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| 特記事項 |
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ICC/IF
Use a concentration of 5 µg/ml. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa). |
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IP
Use a concentration of 5 µg/ml. |
ターゲット情報
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機能
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. -
関連疾患
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. -
配列類似性
Belongs to the PRKRA family.
Contains 3 DRBM (double-stranded RNA-binding) domains. -
ドメイン
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. -
翻訳後修飾
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. -
細胞内局在
Cytoplasm > perinuclear region. - Information by UniProt
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参照データベース
- Entrez Gene: 282875 Cow
- Entrez Gene: 8575 Human
- Entrez Gene: 23992 Mouse
- Entrez Gene: 311130 Rat
- Omim: 603424 Human
- SwissProt: Q2HJ92 Cow
- SwissProt: O75569 Human
- SwissProt: Q9WTX2 Mouse
see all -
別名
- DYT16 antibody
- HSD14 antibody
- Interferon inducible double stranded RNA dependent protein kinase activator A antibody
see all
画像
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Immunocytochemistry/ Immunofluorescence - Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967)
ab31967 staining PACT (PKR activating protein) / PRKRA in Hek293 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 overnight at 4°C with ab31967 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in green) and ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967)All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PRKRA knockout HEK-293T cell lysate
Lane 3 : K-562 cell lysate
Lane 4 : HepG2 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: 34 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab31967 observed at 36 kDa. Red - loading control ab8245 observed at 36 kDa.
ab31967 Anti-PACT (PKR activating protein) / PRKRA antibody was shown to specifically react with PACT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266806 (knockout cell lysate ab258141) was used. Wild-type and PACT knockout samples were subjected to SDS-PAGE. ab31967 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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Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967)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 1/10000 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 1/10000 dilution for 1 h at room temperature before imaging. -
Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967)All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human PACT (PKR activating protein) / PRKRA peptide (ab30768) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Additional bands at: 60 kDa (possible cross reactivity, but this band is not blocked) -
Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967)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
All lanes : 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. -
Immunoprecipitation - Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967)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
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
参考文献 (4)
ab31967 は 4 報の論文で使用されています。
- Kuo YP et al. A novel antisense RNA ASPACT confers multi-level suppression of PACT and associated signalling. RNA Biol 16:1263-1274 (2019). PubMed: 31135270
- Lui PY et al. PACT Facilitates RNA-Induced Activation of MDA5 by Promoting MDA5 Oligomerization. J Immunol 199:1846-1855 (2017). PubMed: 28760879
- Warner MJ et al. S6K2-mediated regulation of TRBP as a determinant of miRNA expression in human primary lymphatic endothelial cells. Nucleic Acids Res 44:9942-9955 (2016). WB ; Human . PubMed: 27407113
- 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 . PubMed: 21464971