Anti chk1

Manufactured by Abcam

Anti-CHK1 is a primary antibody that recognizes the Checkpoint kinase 1 (CHK1) protein. CHK1 is a serine/threonine-protein kinase that plays a crucial role in the cellular response to DNA damage and is involved in cell cycle regulation. This antibody can be used in various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry, to detect and study the expression and localization of the CHK1 protein.

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Mouse anti-Chk1 (2 citations)

3 protocols using anti chk1

Immunoblotting Analysis of DNA Damage Response Proteins

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Cells were plated in a six-well plate at 2 × 10⁵ cells/well overnight. Cells were transfected with 5 nM corresponding siRNA for 72 h. Cells were then lysed in M-PER (Thermo Scientific) buffer containing a protease inhibitor cocktail (Sigma-Aldrich) and a phosphatase inhibitor cocktail (Sigma-Aldrich).
Forty micrograms of protein were subject to immunoblotting analysis. Proteins were separated by NuPAGE 4–12% Bis-Tris gel (Invitrogen) and transferred to PVDF membrane by iBlot 2 Transfer Stacks (Thermo Fisher). Anti-CHK1 (1:5000 dilution, Abcam), anti-RRM1, anti-RRM2, Phospho-Histone H2AX (1:1000 dilution, Cell Signaling) and anti-GAPDH (1:1000 dilution, Santa Cruz) antibodies were used to detect proteins. Proteins were visualized with Alexa Fluor 647 or 488 conjugated secondary antibodies (Invitrogen) using an Azure c400 Imaging System (Azure Biosystems).

Simonenko V., Lu X., Roesch E., Mutisya D., Shao C., Sun Q., Patterson-Orazem A., McNair M., Shanmuganathan A., Lu P, & Evans D.M. (2020). A novel siRNA–gemcitabine construct as a potential therapeutic for treatment of pancreatic cancer. NAR Cancer, 2(3), zcaa016.

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Investigating DNA Damage Response Signaling

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N6-isopentenyladenosine (iPA) (Sigma-Aldrich, St. Louis, MO) was dissolved in DMSO and added to cell cultures at the indicated concentration. For Western blot analysis the following antibodies were used: anti-RAD51, anti-pCHK1 (S345), rabbit anti-pCHK2 (T68), anti-p-ATM (S1981), anti-p-BRCA1 (S1524), anti-p-ATR (S428), anti-p-AKT (S473), anti-PARP, anti- p-JAK2 (Tyr 1007/1008), anti-JAK2, anti-NF-κB p65 (D14E12), and anti-Caspase-3 were purchased from Cell Signaling Technology (Danvers, MA), anti-CHK2, anti-STAT5 a/b, anti-H2AX, anti-γ-H2AX (Ser139), anti-β-actin, anti-BRCA1, anti-p-STAT5a/b (Tyr 694/699), anti-p-p38 (Tyr182) were purchased from Santa Cruz Biotechnology (Dallas, TX), anti-CHK1 from Abcam (Cambridge, UK), anti-BCL-2 and anti-p38 from Sigma-Aldrich Inc. (St Luis, MO). For fluorescence microscopy anti-RAD51 (Cell Signaling Technology, Danvers, MA), anti-γ-H2AX (Santa Cruz Biotechnology Dallas, TX) and Alexa Fluor 488 donkey anti-rabbit IgG (Jackson ImmunoResearch, Cambridge, UK) and DyLight 594 goat anti-mouse IgG (Abcam, Cambridge, UK) were used. STAT5a/b-siRNA and scramble-siRNA were purchased from Santa Cruz Biotechnology (Dallas, TX).

Navarra G., Pagano C., Pacelli R., Crescenzi E., Longobardi E., Gazzerro P., Fiore D., Pastorino O., Pentimalli F., Laezza C, & Bifulco M. (2020). N6-Isopentenyladenosine Enhances the Radiosensitivity of Glioblastoma Cells by Inhibiting the Homologous Recombination Repair Protein RAD51 Expression. Frontiers in Oncology, 9, 1498.

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Western Blot Analyses of Epigenetic Regulators

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Standard western immunoblotting procedures were used with the following antibodies: anti-H3K36me1 (Abcam), anti-H3K36me2 (Abcam), anti-H3K36me3 (Abcam), anti-SETD2 (Santa Cruz Biotechnology), anti-hMSH6 (Santa Cruz Biotechnology), anti-histone 3.3, anti-histone 3, anti-SKP2 (Santa Cruz Biotechnology), anti-biotin (Santa Cruz Biotechnology), anti-Ub (Santa Cruz Biotechnology), anti-P300 (Abcam), anti-RNA Pol II (Abcam), anti-CyclinE (Santa Cruz Biotechnology), anti-CDK4 (Santa Cruz Biotechnology), anti-CyclinD1 (Abcam), anti-PCNA (Abcam), anti-ppRB (Abcam), anti-E2F1 (Abcam), anti-P18 (Abcam), anti-P21/WAF1/Cip1 (Santa Cruz Biotechnology), anti-PKM2, anti-c-Myc (Santa Cruz Biotechnology), anti-Chk1 (Abcam), anti-P62 (Abcam), anti-KDM4A (Abcam), and anti-β-actin (Abcam).
The primers were as follows: P62: P1, 5′-GCAGTATCCCAAGTTCAATT-3′; P2, 5′-TGGGAACAGGTGGTGGAGGA-3′; P62 promoter: P1, 5′-GATCATTCACACCTGTGGAC-3′; P2, 5′-GGACGAGTGGTCACCCTCTG-3′; pre-miR-675: P1, 5′-CCCAGGGTCTGGTGCGGAGA-3′; P2, 5′-CCCAGGGGCTGAGCGGTGAG-3; mature mi675: P1, 5′-TGGTGCGGAGAGGGCCACAGUG-3′; U6 primer: P1, 5′-GCTTCGGCAGCACATATACT-3′; P2, 5′-GGAACGCTTCACGAATTTGC-3′; and β-actin: P1, 5′-CTTCCTTCCTGGGCATGGAG-3′; P2, 5′-TGGAGGGGCCGGACTGGTCA-3′.

Lu Y., Song S., Jiang X., Meng Q., Wang C., Li X., Yang Y., Xin X., Zheng Q., Wang L., Pu H., Gui X., Li T, & Lu D. (2018). miR675 Accelerates Malignant Transformation of Mesenchymal Stem Cells by Blocking DNA Mismatch Repair. Molecular Therapy. Nucleic Acids, 14, 171-183.

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