参阅全部 CDK1 一抗
描述小鼠单克隆抗体[A17] to CDK1
特异性The antibody inhibits the activation of p34cdc2 kinase by cyclins. A study published in the Biotechniques journal suggests that ab18 cross-reacts with Cep152 protein. This should not be a problem for simple western blot experiments because of the difference in molecular weight (MW) between Cdk1 and Cep152. (Cdk1 has a predicted MW of 34-kDa. Isoforms 1-4 of Cep152 have predicted MW of 147, 158, 189 and 196-kDa respectively.) However, this could potentially cause misleading results in Immunofluorescence and pull down experiments. Abcam welcomes customer feedback.
经测试应用适用于: ICC/IF, Flow Cyt, WB, IP, ELISA, IHC-FoFr, IHC-P, RIA, IHC-Frmore details
种属反应性与反应: Mouse, Rat, Chicken, Human, Xenopus laevis
Recombinant fragment corresponding to Xenopus laevis CDK1 aa 50 to the C-terminus (C terminal).
Database link: P24033
表位The epitope is thought to be residues 220-227 of mouse cdc2, LGTPNNEV.
- WB: HeLa, Jurkat, MCF7, A431, RAW 264.7 and NIH/3T3 whole cell lysate. IHC-P: Human skin. Human lung cancer tissue. FC: MCF7 cells.
存放说明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.
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
纯度Immunogen affinity purified
Our Abpromise guarantee covers the use of ab18 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|Flow Cyt||Use 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.|
|IP||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|IHC-FoFr||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|RIA||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
功能Plays a key role in the control of the eukaryotic cell cycle. It is required in higher cells for entry into S-phase and mitosis. p34 is a component of the kinase complex that phosphorylates the repetitive C-terminus of RNA polymerase II.
组织特异性Isoform 2 is found in breast cancer tissues.
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
形式CDK1 can be located to the Nucleus, cytoplasm and Mithocondria. It's cytoplasmic during interphase and reversibly translocated from cytoplasm to the nucleus when phosphorilated before G2-M transition when associated with cyclin-B1. Accumulates in mitochondria in G2-arrested cells upon DNA-damage.
- Cdc 2 antibody
- Cdc2 antibody
- CDC28A antibody
All lanes : Anti-CDK1 antibody [A17] (ab18) at 5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab97046) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutes
This blot was produced using a 4-12% 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 2% Bovine Serum Albumin before being incubated with ab18 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.
Ab18 staining human skin. Staining is localized to the cytoplasm and nucleus.
Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Overlay histogram showing MCF7 cells stained with ab18 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab18, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunohistochemical analysis of Human lung cancer tissue, staining CDK1 with ab18 at 1/100 dilution.
This product has been referenced in:
- Shu F et al. Stathmin gene silencing suppresses proliferation, migration and invasion of gastric cancer cells via AKT/sCLU and STAT3 signaling. Int J Oncol 54:1086-1098 (2019). Read more (PubMed: 30628664) »
- Cho CH et al. Gadd45b Acts as Neuroprotective Effector in Global Ischemia-Induced Neuronal Death. Int Neurourol J 23:S11-21 (2019). Read more (PubMed: 30832463) »