重组Anti-Cdk2抗体[E304] (ab32147)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E304] to Cdk2
- Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Cdk2抗体[E304]
参阅全部 CDK2 一抗 -
描述
兔单克隆抗体[E304] to Cdk2 -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Cdk2 aa 250 to the C-terminus (C terminal). The exact sequence is proprietary.
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表位
The epitope is within the C-terminus of human Cdk2 -
阳性对照
- HeLa cells HeLa whole cell lysate (ab150035).
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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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E304 -
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ab32147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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ICC/IF | (3) |
1/200.
For unpurified use at 1/100. |
IP |
1/40.
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WB | (5) |
1/1000 - 1/10000. Detects a band of approximately 33 kDa (predicted molecular weight: 34 kDa).
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IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/80.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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ICC/IF
1/200. For unpurified use at 1/100. |
IP
1/40. |
WB
1/1000 - 1/10000. Detects a band of approximately 33 kDa (predicted molecular weight: 34 kDa). |
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/80. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Involved in the control of the cell cycle. Interacts with cyclins A, B1, B3, D, or E. Activity of CDK2 is maximal during S phase and G2. -
序列相似性
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. - Information by UniProt
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数据库链接
- Entrez Gene: 1017 Human
- Entrez Gene: 12566 Mouse
- Entrez Gene: 362817 Rat
- Omim: 116953 Human
- SwissProt: P24941 Human
- SwissProt: P97377 Mouse
- SwissProt: Q63699 Rat
- Unigene: 19192 Human
see all -
别名
- Cdc2 related protein kinase antibody
- cdc2-related protein kinase antibody
- CDC28 antibody
see all
图片
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All lanes : Anti-Cdk2 antibody [E304] (ab32147) at 1/2000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : CDK2 knockout MCF7 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDaWestern blot: Anti-CDK2 antibody [E304] (ab32147) staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32147 was shown to bind specifically to CDK2. A band was observed at 34 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CDK2 knockout cell line ab282628. To generate this image, wild-type and CDK2 knockout MCF7 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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Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: CDK2 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Green signal from target – ab32147 observed at 34 kDa
Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signalab32147 was shown to specifically react with CDK2 when CDK2 knockout samples were used. Wild-type and CDK2 knockout samples were subjected to SDS-PAGE. ab32147 and ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4ºC. 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 1/10 000 dilution for 1 h at room temperature before imaging.
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ab32147 staining Cdk2 in the HeLa cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/200). ab150078 (1/500) an Alexa Fluor® 555-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.
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ab32147 staining Cdk2 in human tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/50). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Cdk2 with purified ab32147 at 1/80 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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ab32147 (purified) at 1/40 immunoprecipitating Cdk2 from HeLa cells(Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Lanes 1: Wild-type HAP1 cell lysate (20 µg)
Lanes 2: CDK2 knockout HAP1 cell lysate (20 µg)
Lanes 1 - 2: Merged signal (red and green). Green - ab32147 observed at 34 kDa. Red - loading control, ab8226, observed at 42 kDa or ab18058, observed at 130 kDa.This western blot image is a comparison between ab32147 and a competitor's top cited rabbit polyclonal antibody.
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ab32147 staining Cdk2 in human squamous cell carcinoma of cervix tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/50). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.
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All lanes : Anti-Cdk2 antibody [E304] (ab32147) at 1/5000 dilution
Lane 1 : C6 cell lysate
Lane 2 : PC-12 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 34 kDa -
Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution + NIH/3T3 cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 34 kDa -
All lanes : Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : Hela cell lysate
Lane 3 : K562 cell lysate
Lane 4 : 293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 34 kDa -
All lanes : Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution (unpurified)
Lane 1 : Human osteosarcoma whole cell lysate - control, non-targeting siRNA
Lane 2 : Human osteosarcoma whole cell lysate - siRNA for CDK2
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 2 seconds
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All lanes : Anti-Cdk2 antibody [E304] (ab32147) at 1/1000 dilution (unpurified)
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 10 µg
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 4 minutesThis blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab32147 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
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 (214)
ab32147 被引用在 214 文献中.
- Cao C et al. Propofol prevents the aggressive progression of oral squamous cell carcinoma via regulating circ_0005623/miR-195-5p/HOXB7 axis. Biotechnol Appl Biochem 69:1015-1028 (2022). PubMed: 33894003
- Heo SY et al. Dieckol induces cell cycle arrest by down-regulating CDK2/cyclin E in response to p21/p53 activation in human tracheal fibroblasts. Cell Biochem Funct 40:71-78 (2022). PubMed: 34708431
- Liu Y et al. Diane-35 and Metformin Induce Autophagy and Apoptosis in Polycystic Ovary Syndrome Women with Early-Stage Endometrial Carcinoma. Genes (Basel) 13:N/A (2022). PubMed: 35052471
- Zheng X et al. TAK1 accelerates transplant arteriosclerosis in rat aortic allografts by inducing autophagy in vascular smooth muscle cells. Atherosclerosis 343:10-19 (2022). PubMed: 35078016
- Luo Y et al. DTL Is a Prognostic Biomarker and Promotes Bladder Cancer Progression through Regulating the AKT/mTOR axis. Oxid Med Cell Longev 2022:3369858 (2022). PubMed: 35103094