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    products/panels/cell-cycle-pcdkphh3actin-wb-cocktail-ab136810.pdf

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Cell Cycle (pCdk/pHH3/Actin) WB Cocktail (ab136810)

  • Datasheet
  • SDS
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Western blot – ab136810 Cell Cycle Western Blot Cocktail
  • Western blot – ab136810 Cell Cycle Western Blot Cocktail
  • Western blot – ab136810 Cell Cycle Western Blot Cocktail

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Overview

  • Product name

    Cell Cycle (pCdk/pHH3/Actin) WB Cocktail
  • Species reactivity

    Reacts with: Mouse, Human
  • Product overview

    The Cell Cycle western blot cocktail (ab136810) is designed to provide a rapid assessment of cell cycle distribution based on molecular markers for the G1/S and M phases of the cell cycle. This is a mixture of three specific rabbit monoclonal primary antibodies targeting phospho-cdk2 Tyr15, phospho Histone H3 Ser10 and beta-actin. Cyclin-dependent kinase 2 (Cdk2) is maintained in an inactive state in G1/S by inhibitory phosphorylation on Tyr15. Histone H3 is phosphorylated at Ser10 when chromosomes condense during mitosis. Hence, elevated phospho-cdk2 Tyr15 indicates G1/S arrested cells and elevated phospho Histone H3 Ser10 indicates M-phase arrested cells when compared to asynchronous cycling control cells. An anti- Actin antibody is included as a loading control. These three readouts are easily resolved by western blot given their different molecular weights.


    The recommended dilution for this cocktail is 1:250.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 200 µl
    250X Cell Cycle WB Cocktail 1 x 200µl
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Antibody Cocktails
    • Apoptosis & Cell Cycle

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab136810 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
Use at an assay dependent concentration.

1/250 dilution for primary antibody cocktail

Suggested dilution buffer: 5% milk/PBS

Notes
WB
Use at an assay dependent concentration.

1/250 dilution for primary antibody cocktail

Suggested dilution buffer: 5% milk/PBS

Images

  • Western blot – ab136810 Cell Cycle Western Blot Cocktail
    Western blot – ab136810 Cell Cycle Western Blot Cocktail
    Western blot – ab136810 Cell Cycle Western Blot Cocktail Cdk2 pTyr15 and Histone H3 pSer10 bands are present in asynchronous cells. Cdk2 pTyr15 is elevated in G1/S arrested cells. Conversely, Histone H3 pSer10 is elevated in G2/M arrested cells. Primary antibody: All lanes Cell 250X Cell Cycle WB Cocktail diluted to 1X in 4% milk/PBS. Lane 1: HeLa lysate; Untreated, asynchronous cells Lane 2: HeLa lysate; G1/S arrested cells (thymidine treatment) Lane 3: HeLa lysate; G2/M arrested cells (sequential thymidine and nocodazole treatments) All lysates at 15 µg per lane. Lysates are ab136811 HeLa Cell Cycle Lysates Secondary antibody: All lanes anti-rabbit-HRP. Predicted band size: 42, 33, 17 kDa.
  • Western blot – ab136810 Cell Cycle Western Blot Cocktail
    Western blot – ab136810 Cell Cycle Western Blot Cocktail
    Western blot – ab136810 Cell Cycle Western Blot Cocktail Hydroxyurea, Camptothecin and Etoposide treatments cause a G1/S-like arrest as judged by the increase in Cdk2 pTyr15 and decrease in Histone H3 pSer10 bands relative to control. In contrast, Paclitaxel causes a G2/M like arrest judged by the decreased Cdk2 pTyr15 and increased Histone H3 pSer10 bands. Primary antibody: All lanes Cell 250X Cell Cycle WB Cocktail diluted to 1X in 4% milk/PBS. Lane 1: HeLa lysate; Control treatment Lane 2: HeLa lysate; 5 mM Hydroxyurea, 24 h treatment Lane 3: HeLa lysate; 5 µM Camptothecin, 24 h treatment Lane 4: HeLa lysate; 1.5 µM Etoposide, 24 h treatment Lane 5: HeLa lysate; 2000 nM Paclitaxel, 24 h treatment All lysates at 15 µg per lane. Secondary antibody: All lanes anti-rabbit-HRP. Predicted band size: 42, 33, 17 kDa.
  • Western blot – ab136810 Cell Cycle Western Blot Cocktail
    Western blot – ab136810 Cell Cycle Western Blot Cocktail
    Western blot – ab136810 Cell Cycle Western Blot Cocktail Cdk2 pTyr15 intensity decreases and Histone H3 pSer10 intensity increases as a function of Paclitaxel concentration. Equivalent protein loading is indicated by the equal actin intensity across samples. Primary antibody: All lanes Cell 250X Cell Cycle WB Cocktail diluted to 1X in 4% milk/PBS. Lane 1: HeLa lysate; Control treatment Lane 2: HeLa lysate; 0.25 nM Paclitaxel, 24 h treatment Lane 3: HeLa lysate; 5 nM Paclitaxel, 24 h treatment Lane 4: HeLa lysate; 100 nM Paclitaxel, 24 h treatment Lane 5: HeLa lysate; 2000 nM Paclitaxel, 24 h treatment All lysates at 15 µg per lane. Secondary antibody: All lanes anti-rabbit-HRP. Predicted band size: 42, 33, 17 kDa.

Protocols

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.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (5)

Publishing research using ab136810? Please let us know so that we can cite the reference in this datasheet.

ab136810 has been referenced in 5 publications.

  • Le T  et al. Chimera RNA interference knockdown of ?-synuclein in human cortical astrocytes results in mitotic catastrophe. Neural Regen Res 15:1894-1902 (2020). PubMed: 32246638
  • Vanoni S  et al. Identification of anoctamin 1 (ANO1) as a key driver of esophageal epithelial proliferation in eosinophilic esophagitis. J Allergy Clin Immunol 145:239-254.e2 (2020). PubMed: 31647967
  • Kuo KL  et al. The Deubiquitinating Enzyme Inhibitor PR-619 Enhances the Cytotoxicity of Cisplatin via the Suppression of Anti-Apoptotic Bcl-2 Protein: In Vitro and In Vivo Study. Cells 8:N/A (2019). PubMed: 31627336
  • Tong WY  et al. Delivery of siRNA in vitro and in vivo using PEI-capped porous silicon nanoparticles to silence MRP1 and inhibit proliferation in glioblastoma. J Nanobiotechnology 16:38 (2018). PubMed: 29653579
  • Planells-Palop V  et al. Human germ/stem cell-specific gene TEX19 influences cancer cell proliferation and cancer prognosis. Mol Cancer 16:84 (2017). WB ; Human . PubMed: 28446200

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