重组Anti-Chk1抗体[EP691Y] (ab40866)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP691Y] to Chk1
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Chk1抗体[EP691Y]
参阅全部 Chk1 一抗 -
描述
兔单克隆抗体[EP691Y] to Chk1 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Chk1 aa 450-550 (C terminal). The exact sequence is proprietary.
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阳性对照
- HeLa cells lysate and human breast carcinoma.
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常规说明
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP691Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab40866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 |
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Flow Cyt (Intra) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/100 - 1/250.
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WB | (5) |
1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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说明 |
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Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/250. |
WB
1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
靶标
- Information by UniProt
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数据库链接
- Entrez Gene: 1111 Human
- Omim: 603078 Human
- SwissProt: O14757 Human
- Unigene: 24529 Human
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别名
- C85740 antibody
- Cell cycle checkpoint kinase antibody
- Checkpoint , S. pombe, homolog of, 1 antibody
see all
图片
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All lanes : Anti-Chk1 antibody [EP691Y] (ab40866) at 1/10000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : CHEK1 knockout A549 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 57 kDa why is the actual band size different from the predicted?Anti-CHEK1 antibody [EP691Y] (ab40866) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40866 was shown to bind specifically to CHEK1. A band was observed at 57 kDa in wild-type A549 cell lysates with a reduction in signal observed at this size in CHEK1 heterozygous knockout cell line. To generate this image, wild-type and CHEK1 heterozygous 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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Anti-Chk1 antibody [EP691Y] (ab40866) at 1/10000 dilution + HeLa cell lysate at 10 µg
Predicted band size: 54 kDa
Observed band size: 54 kDa -
Immunofluorescence staining of MCF7 cells with purified ab40866 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.
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Overlay histogram showing HeLa cells stained with ab40866 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40866, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chk1 antibody [EP691Y] (ab40866)
Immunohistochemical analysis of Chk1 expression in paraffin-embedded human breast carcinoma using ab40866 at a 1:250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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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SDS download
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Datasheet download
文献 (33)
ab40866 被引用在 33 文献中.
- Zhou F et al. The role of USP51 in attenuating chemosensitivity of lung cancer cells to cisplatin by regulating DNA damage response. Biotechnol Appl Biochem 70:634-644 (2023). PubMed: 35856403
- Zhong NS et al. HELQ suppresses migration and proliferation of non-small cell lung cancer cells by repairing DNA damage and inducing necrosis. Cell Biol Int 47:188-200 (2023). PubMed: 36183369
- Bruyer A et al. Combined inhibition of Wee1 and Chk1 as a therapeutic strategy in multiple myeloma. Front Oncol 13:1271847 (2023). PubMed: 38125947
- Wang T et al. TOP2A promotes proliferation and metastasis of hepatocellular carcinoma regulated by miR-144-3p. J Cancer 13:589-601 (2022). PubMed: 35069905
- Wang L et al. Long Noncoding RNA Solute Carrier Family 25 Member 21 Antisense RNA 1 Inhibits Cell Malignant Behaviors and Enhances Radiosensitivity of Gastric Cancer Cells by Upregulating Synuclein Gamma Expression. Tohoku J Exp Med 257:225-239 (2022). PubMed: 35444105