Anti-CDK1 抗体 [A17] (ab18)

Reviews (2)Q&A (8)References (58)

製品の概要

  • 製品名

    Anti-CDK1 antibody [A17]
    CDK1 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [A17] to CDK1

  • 由来種

    Mouse

  • 特異性

    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.

  • アプリケーション

    適用あり: ICC/IF, Flow Cyt, WB, IP, ELISA, IHC-FoFr, IHC-P, RIA, IHC-Frmore details

  • 種交差性

    交差種: Mouse, Rat, Chicken, Human, Xenopus laevis

  • 免疫原

    Recombinant fragment corresponding to Xenopus laevis CDK1 aa 50 to the C-terminus (C terminal).
    Database link: P24033

  • エピトープ

    The epitope is thought to be residues 220-227 of mouse cdc2, LGTPNNEV.

  • ポジティブ･コントロール

    • 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.

  • 特記事項

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    Isoform 2 is found in breast cancer tissues.

  • 配列類似性

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
    Contains 1 protein kinase domain.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    see all

  • 製品の状態

    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.

  • 別名

    • Cdc 2 antibody
    • Cdc2 antibody
    • CDC28A antibody
    • CDK 1 antibody
    • CDK1 antibody
    • CDK1_HUMAN antibody
    • CDKN1 antibody
    • CELL CYCLE CONTROLLER CDC2 antibody
    • Cell division control protein 2 antibody
    • Cell division control protein 2 homolog antibody
    • Cell division cycle 2 G1 to S and G2 to M antibody
    • Cell division protein kinase 1 antibody
    • Cell Divsion Cycle 2 Protein antibody
    • Cyclin Dependent Kinase 1 antibody
    • Cyclin-dependent kinase 1 antibody
    • DKFZp686L20222 antibody
    • MGC111195 antibody
    • p34 Cdk1 antibody
    • p34 protein kinase antibody
    • P34CDC2 antibody
    see all

画像

  • Western blot - Anti-CDK1 antibody [A17] (ab18)
    Western blot - Anti-CDK1 antibody [A17] (ab18)
    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.

    Secondary
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 antibody [A17] (ab18)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 antibody [A17] (ab18)
    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.
  • Flow Cytometry - Anti-CDK1 antibody [A17] (ab18)
    Flow Cytometry - Anti-CDK1 antibody [A17] (ab18)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 antibody [A17] (ab18)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 antibody [A17] (ab18)Image from Zhang C et al., PLoS One. 2011;6(8):e23849. Epub 2011 Aug 24. Fig 3.; doi:10.1371/journal.pone.0023849; August 24, 2011, PLoS ONE 6(8): e23849.
    Immunohistochemical analysis of Human lung cancer tissue, staining CDK1 with ab18 at 1/100 dilution.

参考文献

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) »
See all 58 Publications for this product

ab18 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

レビューと Q&A

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1-10 of 10 Abreviews or Q&A

Question

elen Dank für die Zusendung des CdK1(ab18)-Ersatzrörchens
. Der Antikörper kam leider heute erst an und
der Kühlakku war nicht mehr kühl. Ich hoffe, der AK hat dadurch keinen
Schaden genommen, werde ihn aber erstmal testen.

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Answer

Der Antikörper sollte dies eigentlich unbeschadet überstanden haben. Wir versenden unsere Antikörper so z.B. auch nach Australien oder Thailand, und da kann es schon einmal zu Transitzeiten von einer Woche kommen, und bisher haben wir noch keine erhöhten Beschwerderaten für die so versandten Produkte.

Die meisten Antikörper sind für längere Zeit bei Raumtemperatur stabil. Versuche bei Abcam mit Antikörpern, die über einen längeren Zeitraum bei Raumtemperatur gelagert wurden, haben gezeigt, dass die Aktivität erst nach zwei Monaten nachlässt. Dennoch verschicken wir Antikörper als Vorsichtsmaßnahme mit Kühlpaketen, die die Umgebungstemperatur für 48 Stunden bei circa 4°C halten. Dieses verbessert auch die Haltbarkeit der Antikörper und verringert das rapide Auf- und Abkühlen der Antikörper.


Falls es denoch zu Problemen kommen sollte, melden Sie sich bitte uns.


Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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Question

Sehr geehrte Damen und Herren,
ich bitte um technische Hilfestellung zu folgendem Antikörper:
Anti-CDK1 antibody [A17] (ab18), Mouse monoclonal
Ich möchte damit CdK1 in Pancreas Cryoschnitten, wahlweise auch
Paraffinschnitten nachweisen, und parallel dazu Insulin (Abcam, Guinea
Pig). Als Sekundär-AK verwende ich Goat Anti-Mouse-Alexa488 oder-568
bzw. Goat Anti-Guinea-Pig-Alexa568 oder -488. Zellkernfärbung mit DAPI.
Bisher habe ich noch kein spezifisches Signal erhalten. Folgende
Versuche habe ich durchgeführt:
1. CdK1 in Wildtyp Pancreas Cryoschnitten
- frisches Pancreas-Gewebe wurde vor Sucrosereihe und dem einbetten in
Tissue-TEK über Nacht bei 4°C in 4% PFA fixiert
- Cryoschnitte 12um, 30min trocknen, storage -20°C 7 Tage
- Schnitte 30min bei RT trocknen
- Fixation: 15min 4% PFA bei RT
- Blocking und Permeab.: 30min 0.3% Triton in 16% Goat Serum NaPO4
Puffer pH 7.4
- Primary AB über Nacht in Blocking Puffer 4°C: Anti CdK1 1:50 bis 1:200
- Secondary AB 1:200 1h RT in Blocking Puffer (CdK1: Alexa488,
Insulin: Alexa568)
- Ergebnis: unspezifische Färbung auch in Negativ-Kontrolle
2. CdK1 in WT Pancreas Paraffinschnitten
- Gewebe-Fixierung in 4% PFA über Nacht 4°C
- 5um-Schnitte
- Deparaffinisierung in Xylen und Ethanolreihe
- AG-Retrieval: 40min Microwave Boiling in Citratpuffer pH 6.0, 30min abkühlen
- Blocking und Permeab.: 45min RT in 0.2% Saponin 16% Goat Serum NaPO4
Puffer pH 7.4
- Primary AB über Nacht in Blocking Puffer 4°C: Anti CdK1 1:25 bis 1:100
- Secondary AB 1:200 1h RT in Blocking Puffer (CdK1: Alexa488,
Insulin: Alexa568)
-Ergebnis: unspezifische Färbung auch in Negativ-Kontrolle
3. CdK1 in WT Pancreas Paraffinschnitten mit Thermo Scientific Nuclear
Staining Kit
- Paraffinschnitte, Vorrbehandlung und Ag-Retrieval wie bei 2.
- CdK1: Alexa488, Insulin: Alexa568
- Positiv-Kontrolle: BRDU: Alexa546
- siehe Anhang (2012-03-21 CdK1 Test TS Nuclear Staining Kit)
4. CdK1 in WT Dünndarm Paraffinschnitten mit Thermo Scientific Nuclear
Staining Kit (als Positivkontrolle wegen hoher Proliferation im
Dünndarmgewebe)
- Procedure wie bei 3.
- siehe Anhang (2012-03-29 CdK1 (Abcam) TS Nuclear Staining Kit on
Small Intestines)
5. CdK1 in MIN6 Cells: Vergleich Normal CdK1 und CdK1 Overexpression
- Protokoll A: 20min 4% PFA 4°C, Saponin Procedure wie bei 2., Washing
mit Low und High Salt Buffer
- Protokoll B: Abreview
https://www.abcam.com/index.html?datasheet=18&tab=abreviews&intabreviewid=20556
- CdK1: Alexa488
- Kontrollen: Anti-HIS als Positiv-Kontrolle, da das CdK1-Konstrukt
für die Transfektion auch einen HIS-Tag besitzt; CyclinB;
Negativ-Kontrolle: kein Primary AB
- Ergebnis: kein Signal bei CdK1, spezifische Färbung bei HIS, Färbung
bei CyclinB; bessere Färbeergebnisse bei den Kontrollen mit Protokoll B
-siehe Anhang (2012-04-02 CdK1 Test MIN6)
Leider habe ich auch bei Versuch Nr. 5 kein Signal für CdK1 erhalten,
so wie es in dem Abreview-Protokoll für HeLa-Zellen abgebildet ist.
Ich bitte Sie um Ratschläge, wie ich doch noch zu einem Färbeergebnis
kommen kann. Da dies nun schon der dritte Anti-CdK1-AK ist, den ich
erfolglos teste, gehen mir die Ideen aus. Aber irgendwie müssen die
Bilder von CdK1-positiven Zellen und Geweben ja zustande kommen.
Ich freue mich, wenn Sie mir den entscheidenden Hinweis geben können.
Vielen Dank.
Mit freundlichen Grüßen

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Answer

Vielen Dank für IhreAnfrage und all dieInformationen.

Es tut mir leid zu hören, dass dieses Röhrchen vonab18 keine guten Ergebnisse liefert. Ich stimme Ihnen zu, dass kein spezifisches Signal erkannt werden kann in all den von Ihnen durchgeführten Experimenten. Deshalb denke ich, dass Sie ein schlechtes Röhrchen erhalten haben.

Der hohe Hintergrund ist sicherlich auf den Sekundären Antikörper zurückzuführen, da auch die Negativkontrolle ohne primären Antikörper diesen Hintergrund anzeigt.

Ich möchte Ihnen außerdem vorschlagen in zukünftigen Versuchen EDTA, pH 9 als Antigendemaskierungspuffer zu verwenden. Dieser Puffer hat unserem Wissen zufolge meist bessere Ergebnisse zu Folge:

http://www.nordiqc.org/Techniques/Epitope_retrieval.htm

Leider muss meisten auch der Permeabilisierungsschritt optimiert werden. So ist z.B. Saponin reversibel und sollte deshalb in allen Puffern verwendet werden während des Experiments.

Ich weiß, dass Sie viel Zeit für diese Experimente verwendet haben und möchte Ihnen daher ein neues Röhrchen oder einen Gutschein als Ersatz anbieten (vorausgesetzt, das Produkt wurde in den letztensechs Monatengekauft). Dafür benötige ich jedoch ihre ursprüngliche Bestellnummer und die Lotnummeroder einen Kontakt in Ihrer Einkaufsabteilung und das Datum der Bestellung. Würden Sie mir bitte zusammen mit diesen Informationen mitteilen, welche Alternative Sie bevorzugen?

Vielen Dank für Ihre Kooperation und Hilfsbereitschaft. Ich freue mich bald wieder von Ihnen zu hören.

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Question

Thank you very much I will order it as as soon as its availible Kindest regards

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Answer

Thank you for your email. I have just been informed by my colleague that the blocking peptide is out of stock. Therefore they will not be able to publish the datasheet hence we are unable to fulfil your request this time. I have done my best to publish this product without delays however due to non availability of the product this happens to be not possible. I sincerely apologize for the inconvenience this may have caused. I hope this information is nevertheless helpful. Should you have any other question pleasedo not hesitate to contact me.

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Question

I would like to go ahead and order the peptide to antibody ab47329 as a special request. I also need to place an order for ab47329 and also ab18 can you tell me what to put the request down as on my order form and the price please as I was going to put this through as the one order. Thank you

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Answer

Thank you for your email. I have sent request to datasheet team for publishing the datasheet of blocking peptide. They will be able to publish this on Monday next week. I will then send you the catalogue number for ordering. Many thanks for having patience!

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Question

Hello Thank you for getting back to me, yes I would like a peptide for ab47329 made that would be great. I can not order it today as we finish for christmas and are back on the 5th january so I shall be in touch then. Can you tell me what the controls suggested are for and what to do with them Im not sure at the moment Thank you Kindest regards

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Answer

Thank you for contacting us. Isotype controls are the primary antibodies that do not have specificity for any target. These can be used as negative control and are helpful in determining the background staining due to secondary antibodies. By comparing the result of actual primary antibody and isotyope control ab you may determine the target localization easily. No primary control also helps determining the background staining due to secondary antibodies. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

Hello I have been using the Abcam antibody for Cyclin dependent kinase 1 (Ab18) and also the phosphorylated version of this antibody ab47329. I am using these antibodies for a IHC study and was wondering if you have any blocking peptides availible for these anibodies as I need to do antibody specificity experiments. Thank you Kindest regards

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Answer

Thank you for contacting us. We can provide blocking peptide for ab47329 as a special request. Could you please confirm if you are interested in buying this? We unfortunately will not be able to supply the blocking peptide for ab18. I suggest trying alternatives as isotype control or no primary control in experiment. We have isotype control products with catalogue number ab18414, ab91361 etc. https://www.abcam.com/Mouse-IgG2a-kappa-MOPC-173-low-endotoxin-isotype-control-ab18449.html https://www.abcam.com/Mouse-IgG2a-ICIGG2A-Isotype-Control-ab91361.html I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Immunocytochemistry/ Immunofluorescence abreview for Anti-CDK1 antibody [A17]

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Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa cells)
Specification
HeLa cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.05% Triton X 100 in PBS for 5 min
Blocking step
goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

投稿 Sep 29 2010

Western blot abreview for Anti-CDK1 antibody [A17]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - other (HeLa cells)
Loading amount
20 µg
Specification
HeLa cells
Gel Running Conditions
Reduced Denaturing (8% Tris Glycine)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

投稿 Aug 18 2010

Question

I am looking for a loading control for Westerns that is not regulated by androgens. Do you have any reports on whether this or other Cdc's are androgen regulated? Erin

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Answer

We do not have any report that this or any other Cdc's are androgen regulated. We are sorry that we do not have any further information and suggest you refer to the latest literature in the field.

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Question

Hello, I would like to know if this antibody can bind to Rabbit antigens. Ravi

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Answer

This antibody has not yet been tested for cross-reactivity with rabbit.

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