重组Anti-Chk1抗体[E250] (ab32531)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E250] to Chk1
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Chk1抗体[E250]
参阅全部 Chk1 一抗 -
描述
兔单克隆抗体[E250] to Chk1 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IP, ICC/IF, WBmore details
不适用于: IHC -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Chk1 aa 1-100 (N terminal). The exact sequence is proprietary.
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阳性对照
- IP: K-562 whole cell lysate. ICC/IF: HeLa cells. Flow Cyt (intra): K-562 cells. WB: K-562, Hela, NIH/3T3, MEF and PC-12 whole cell lysates
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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.
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 long term. Avoid freeze / thaw cycle. -
存储溶液
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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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E250 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-Chk1 antibody [E250] - BSA and Azide free (ab239821)
- PE Anti-Chk1 antibody [E250] (ab318321)
- APC Anti-Chk1 antibody [E250] (ab318424)
- Alexa Fluor® 488 Anti-Chk1 antibody [E250] (ab318527)
- Alexa Fluor® 647 Anti-Chk1 antibody [E250] (ab318630)
- Alexa Fluor® 594 Anti-Chk1 antibody [E250] (ab318733)
- Alexa Fluor® 555 Anti-Chk1 antibody [E250] (ab318833)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32531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 |
---|---|---|
Flow Cyt (Intra) |
1/30.
|
|
IP |
1/20.
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|
ICC/IF | (2) |
1/50.
|
WB | (2) |
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
|
说明 |
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Flow Cyt (Intra)
1/30. |
IP
1/20. |
ICC/IF
1/50. |
WB
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa). |
靶标
- Information by UniProt
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数据库链接
- Entrez Gene: 1111 Human
- Entrez Gene: 12649 Mouse
- Entrez Gene: 140583 Rat
- Omim: 603078 Human
- SwissProt: O14757 Human
- SwissProt: O35280 Mouse
- SwissProt: Q91ZN7 Rat
- Unigene: 24529 Human
see all -
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 [E250] (ab32531) at 1/1000 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 [E250] (ab32531) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32531 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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All lanes : Anti-Chk1 antibody [E250] (ab32531) at 1/10000 dilution (Purified)
Lane 1 : MEF (Mouse embryonic fibroblast (immortalized)) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 54 kDa
Observed band size: 54 kDaWe are unsure how to define the extra bands.
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All lanes : Purfiied at 1/1000 dilution
Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDaWe are unsure how to define the extra bands.
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Purified ab32531 at 1/20 dilution (1µg) immunoprecipitating Chk1 in K-562 whole cell lysate.
Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
Lane 2 (+): ab32531 + K-562 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32531 in K-562 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 54 kDa -
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Chk1 with purified ab32531 at 1/50 dilution (5.06 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Chk1 with purified ab32531 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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 (12)
ab32531 被引用在 12 文献中.
- Liu C et al. Combining radiation and the ATR inhibitor berzosertib activates STING signaling and enhances immunotherapy via inhibiting SHP1 function in colorectal cancer. Cancer Commun (Lond) 43:435-454 (2023). PubMed: 36855844
- Wang J et al. The deubiquitinase USP28 stabilizes the expression of RecQ family helicases and maintains the viability of triple negative breast cancer cells. J Biol Chem 298:101443 (2022). PubMed: 34822842
- Jin M et al. Circ_0011292 knockdown mitigates progression and drug resistance in PTX-resistant non-small-cell lung cancer cells by regulating miR-433-3p/CHEK1 axis. Thorac Cancer 13:1276-1288 (2022). PubMed: 35348291
- Li N & Wang B Suppressive effects of umbilical cord mesenchymal stem cell-derived exosomal miR-15a-5p on the progression of cholangiocarcinoma by inhibiting CHEK1 expression. Cell Death Discov 8:205 (2022). PubMed: 35428780
- Huang L et al. AZD6738 Inhibits fibrotic response of conjunctival fibroblasts by regulating checkpoint kinase 1/P53 and PI3K/AKT pathways. Front Pharmacol 13:990401 (2022). PubMed: 36204234