重组Anti-DNA PKcs抗体[EPR392] - BSA and Azide free (ab174573)
![Western blot - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.cn/ps/products/174/ab174573/Images/ab174573-515342-anti-prkdc-antibody-epr392-western-blot-wild-type-a549-cell-human.jpg "Western blot - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR392] to DNA PKcs - BSA and Azide free
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-DNA PKcs抗体[EPR392] - BSA and Azide free
参阅全部 DNA PKcs 一抗 -
描述
兔单克隆抗体[EPR392] to DNA PKcs - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IFmore details
不适用于: Flow Cyt,IHC-P or IP -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: K562, Molt4, MCF7, SH-SY5Y, 293T and PC3 cell lysates. ICC/IF: K-562 cells
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常规说明
ab174573 is the carrier-free version of ab133516.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
Constituent: PBS -
无载体
是 -
Concentration information loading... -
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR392 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Conjugation kits
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab174573于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB |
Use at an assay dependent concentration. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa).
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| ICC/IF |
Use at an assay dependent concentration.
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| 说明 |
|---|
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WB
Use at an assay dependent concentration. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa). |
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ICC/IF
Use at an assay dependent concentration. |
靶标
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功能
Serine/threonine-protein kinase that acts as a molecular sensor for DNA damage. Involved in DNA nonhomologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination. Must be bound to DNA to express its catalytic properties. Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step. Required to protect and align broken ends of DNA. May also act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage. Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion. Also involved in modulation of transcription. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX, thereby regulating DNA damage response mechanism. Phosphorylates DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, SRF, XRCC1, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2. Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA. Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D. -
序列相似性
Belongs to the PI3/PI4-kinase family.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 2 HEAT repeats.
Contains 1 PI3K/PI4K domain.
Contains 3 TPR repeats. -
翻译后修饰
Phosphorylated upon DNA damage, probably by ATM or ATR. Autophosphorylated on Thr-2609, Thr-2638 and Thr-2647. Thr-2609 is a DNA damage-inducible phosphorylation site (inducible with ionizing radiation, IR). Autophosphorylation induces a conformational change that leads to remodeling of the DNA-PK complex, requisite for efficient end processing and DNA repair.
S-nitrosylated by GAPDH. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 5591 Human
- Omim: 600899 Human
- SwissProt: P78527 Human
- Unigene: 491682 Human
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别名
- DNA dependent protein kinase catalytic subunit antibody
- DNA PK catalytic subunit antibody
- DNA-dependent protein kinase catalytic subunit antibody
see all
图片
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Western blot - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)All lanes : Anti-DNA PKcs antibody [EPR392] (ab133516) at 1/2000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : PRKDC knockout A549 cell lysate
Lane 3 : K562 cell lysate
Lane 4 : HDLM-2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 469 kDa
Observed band size: 450 kDa why is the actual band size different from the predicted?This WB data was generated using the same anti-DNA PKcs antibody clone, EPR392, in a different buffer formulation (ab133516).
Anti-PRKDC antibody [EPR392] (ab133516) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133516 was shown to bind specifically to PRKDC. A band was observed at 450 kDa in wild-type A549 cell lysates with no signal observed at this size in PRKDC knockout cell line. To generate this image, wild-type and PRKDC knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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![Western blot - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.cn/ps/products/174/ab174573/Images/ab174573-309380-ab133516KOWB.jpg)
Western blot - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)This WB data was generated using the same anti-DNA PKcs antibody clone, EPR392, in a different buffer formulation (cat# ab133516).
Lane 1: Wild type HAP1 whole cell lysate (40 µg)
Lane 2: DNA PKcs knockout HAP1 whole cell lysate (40 µg)
Lane 3: K562 whole cell lysate (20 µg)
Lane 4: SHSY5Y whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab133516 observed at 460 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab133516 was shown to specifically react with DNA PKcs when DNA PKcs knockout samples were used. Wild-type and DNA PKcs knockout samples were subjected to SDS-PAGE. Ab133516 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.cn/ps/products/174/ab174573/Images/ab174573-3-ab133516ICCIF1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)Immunocytochemistry/ Immunofluorescence analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling DNA PKcs with purified ab133516 at 1:200 dilution (9.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
![OI-RD Scanning - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.cn/ps/products/174/ab174573/Images/ab174573-309379-ab133516Affinity.jpg)
OI-RD Scanning - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)This kD data was generated using the same anti-DNA PKcs antibody clone, EPR392, in a different buffer formulation (cat# ab133516).
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KD -
![Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.cn/ps/products/174/ab174573/Images/ab174573-4-benefits-of-recombinant-antibodies.png)
Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)
实验方案
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.
数据表及文件
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Datasheet download
文献 (0)
ab174573 尚未被引用在任何文献中。
