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chk1  (Novus Biologicals)
90
Novus Biologicals chk1
Figure 1. KRCC1 promotes <t>CHK1</t> activation and efficient checkpoint. (A, B) OV90 and U2OS cells were transfected with control siRNA (siCTL) or siRNA targeting KRCC1 (siKRCC1) for 72 h. Cells were stained with DAPI and anti-RPA2 antibody and visualized by fluorescence microscopy (scale bar, 20 m). Percentage of cells with >10 foci is quantitated. The data are from manual scoring of ∼200 cells per condition and from three experiments ± SDs. (C) Lysates from above transfected siCTL and siKRCC1 OV90 and U2OS cells were analyzed by immunoblotting for CHK1-mediated DDR markers. (D) Immunoblotting for CHK1-mediated DDR markers after 72 h KRCC1 silencing in the presence or absence of CPT (1 M for 1 h). (E) KRCC1 interaction with CHK1 and 14-3-3 was evaluated using co-immunoprecipitation in EV or HA-tagged KRCC1 (HA-KRCC1) overexpressed cells treated with or without CPT (3 M, 2 h). (F) CHK1 interaction with KRCC1 was evaluated using co-immunoprecipitation in EV or Halo-tagged KRCC1 (Halo-KRCC1) overexpressed cells treated with CPT (3 M, 2 h) in the presence or absence of ATR inhibitor (ATRi, 5 M, 4 h) and quantification of immunoprecipitated Halo-KRCC1 by densitometry analysis using NIH ImageJ and normalized to their respective CHK1 levels and compared to CPT treatment only, which was set to 1. Experiments were repeated three times. Data represent mean ± SD. Marker (M) and IgG lanes are shown. (G) OV90 cells transfected with EV or HA-KRCC1 were treated with CPT (1 M for 1 h) and released for up to 180 min. Cells were collected at the indicated time points and processed for immunoblotting. The black right-pointing triangle indicates KRCC1. Short exposure (SE) and long exposure (LE) blots for KRCC1 are shown. The right panel depicts quantification of pCHK1-S296 by densitometry analysis using NIH ImageJ, normalized to their respective CHK1 levels and compared to the no treatment control group (NT), which was set to 1. (H) Proposed model of CHK1 activation. Following DNA damage and ATR-mediated phosphorylation of CHK1 at S345, CHK1 associates directly or indirectly with KRCC1 and 14-3-3. We posit that this association induces a conformational change in CHK1 to favor autophosphorylation at S296 and enhance kinase activity toward CDC25A.
Chk1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/chk1/product/Novus Biologicals
Average 90 stars, based on 1 article reviews
chk1 - by Bioz Stars, 2026-04
90/100 stars
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goat polyclonal anti chk1 antibody  (R&D Systems)
92
R&D Systems goat polyclonal anti chk1 antibody
Figure 1. KRCC1 promotes <t>CHK1</t> activation and efficient checkpoint. (A, B) OV90 and U2OS cells were transfected with control siRNA (siCTL) or siRNA targeting KRCC1 (siKRCC1) for 72 h. Cells were stained with DAPI and anti-RPA2 antibody and visualized by fluorescence microscopy (scale bar, 20 m). Percentage of cells with >10 foci is quantitated. The data are from manual scoring of ∼200 cells per condition and from three experiments ± SDs. (C) Lysates from above transfected siCTL and siKRCC1 OV90 and U2OS cells were analyzed by immunoblotting for CHK1-mediated DDR markers. (D) Immunoblotting for CHK1-mediated DDR markers after 72 h KRCC1 silencing in the presence or absence of CPT (1 M for 1 h). (E) KRCC1 interaction with CHK1 and 14-3-3 was evaluated using co-immunoprecipitation in EV or HA-tagged KRCC1 (HA-KRCC1) overexpressed cells treated with or without CPT (3 M, 2 h). (F) CHK1 interaction with KRCC1 was evaluated using co-immunoprecipitation in EV or Halo-tagged KRCC1 (Halo-KRCC1) overexpressed cells treated with CPT (3 M, 2 h) in the presence or absence of ATR inhibitor (ATRi, 5 M, 4 h) and quantification of immunoprecipitated Halo-KRCC1 by densitometry analysis using NIH ImageJ and normalized to their respective CHK1 levels and compared to CPT treatment only, which was set to 1. Experiments were repeated three times. Data represent mean ± SD. Marker (M) and IgG lanes are shown. (G) OV90 cells transfected with EV or HA-KRCC1 were treated with CPT (1 M for 1 h) and released for up to 180 min. Cells were collected at the indicated time points and processed for immunoblotting. The black right-pointing triangle indicates KRCC1. Short exposure (SE) and long exposure (LE) blots for KRCC1 are shown. The right panel depicts quantification of pCHK1-S296 by densitometry analysis using NIH ImageJ, normalized to their respective CHK1 levels and compared to the no treatment control group (NT), which was set to 1. (H) Proposed model of CHK1 activation. Following DNA damage and ATR-mediated phosphorylation of CHK1 at S345, CHK1 associates directly or indirectly with KRCC1 and 14-3-3. We posit that this association induces a conformational change in CHK1 to favor autophosphorylation at S296 and enhance kinase activity toward CDC25A.
Goat Polyclonal Anti Chk1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/goat polyclonal anti chk1 antibody/product/R&D Systems
Average 92 stars, based on 1 article reviews
goat polyclonal anti chk1 antibody - by Bioz Stars, 2026-04
92/100 stars
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Figure 1. KRCC1 promotes CHK1 activation and efficient checkpoint. (A, B) OV90 and U2OS cells were transfected with control siRNA (siCTL) or siRNA targeting KRCC1 (siKRCC1) for 72 h. Cells were stained with DAPI and anti-RPA2 antibody and visualized by fluorescence microscopy (scale bar, 20 m). Percentage of cells with >10 foci is quantitated. The data are from manual scoring of ∼200 cells per condition and from three experiments ± SDs. (C) Lysates from above transfected siCTL and siKRCC1 OV90 and U2OS cells were analyzed by immunoblotting for CHK1-mediated DDR markers. (D) Immunoblotting for CHK1-mediated DDR markers after 72 h KRCC1 silencing in the presence or absence of CPT (1 M for 1 h). (E) KRCC1 interaction with CHK1 and 14-3-3 was evaluated using co-immunoprecipitation in EV or HA-tagged KRCC1 (HA-KRCC1) overexpressed cells treated with or without CPT (3 M, 2 h). (F) CHK1 interaction with KRCC1 was evaluated using co-immunoprecipitation in EV or Halo-tagged KRCC1 (Halo-KRCC1) overexpressed cells treated with CPT (3 M, 2 h) in the presence or absence of ATR inhibitor (ATRi, 5 M, 4 h) and quantification of immunoprecipitated Halo-KRCC1 by densitometry analysis using NIH ImageJ and normalized to their respective CHK1 levels and compared to CPT treatment only, which was set to 1. Experiments were repeated three times. Data represent mean ± SD. Marker (M) and IgG lanes are shown. (G) OV90 cells transfected with EV or HA-KRCC1 were treated with CPT (1 M for 1 h) and released for up to 180 min. Cells were collected at the indicated time points and processed for immunoblotting. The black right-pointing triangle indicates KRCC1. Short exposure (SE) and long exposure (LE) blots for KRCC1 are shown. The right panel depicts quantification of pCHK1-S296 by densitometry analysis using NIH ImageJ, normalized to their respective CHK1 levels and compared to the no treatment control group (NT), which was set to 1. (H) Proposed model of CHK1 activation. Following DNA damage and ATR-mediated phosphorylation of CHK1 at S345, CHK1 associates directly or indirectly with KRCC1 and 14-3-3. We posit that this association induces a conformational change in CHK1 to favor autophosphorylation at S296 and enhance kinase activity toward CDC25A.

Journal: Nucleic acids research

Article Title: KRCC1, a modulator of the DNA damage response.

doi: 10.1093/nar/gkac890

Figure Lengend Snippet: Figure 1. KRCC1 promotes CHK1 activation and efficient checkpoint. (A, B) OV90 and U2OS cells were transfected with control siRNA (siCTL) or siRNA targeting KRCC1 (siKRCC1) for 72 h. Cells were stained with DAPI and anti-RPA2 antibody and visualized by fluorescence microscopy (scale bar, 20 m). Percentage of cells with >10 foci is quantitated. The data are from manual scoring of ∼200 cells per condition and from three experiments ± SDs. (C) Lysates from above transfected siCTL and siKRCC1 OV90 and U2OS cells were analyzed by immunoblotting for CHK1-mediated DDR markers. (D) Immunoblotting for CHK1-mediated DDR markers after 72 h KRCC1 silencing in the presence or absence of CPT (1 M for 1 h). (E) KRCC1 interaction with CHK1 and 14-3-3 was evaluated using co-immunoprecipitation in EV or HA-tagged KRCC1 (HA-KRCC1) overexpressed cells treated with or without CPT (3 M, 2 h). (F) CHK1 interaction with KRCC1 was evaluated using co-immunoprecipitation in EV or Halo-tagged KRCC1 (Halo-KRCC1) overexpressed cells treated with CPT (3 M, 2 h) in the presence or absence of ATR inhibitor (ATRi, 5 M, 4 h) and quantification of immunoprecipitated Halo-KRCC1 by densitometry analysis using NIH ImageJ and normalized to their respective CHK1 levels and compared to CPT treatment only, which was set to 1. Experiments were repeated three times. Data represent mean ± SD. Marker (M) and IgG lanes are shown. (G) OV90 cells transfected with EV or HA-KRCC1 were treated with CPT (1 M for 1 h) and released for up to 180 min. Cells were collected at the indicated time points and processed for immunoblotting. The black right-pointing triangle indicates KRCC1. Short exposure (SE) and long exposure (LE) blots for KRCC1 are shown. The right panel depicts quantification of pCHK1-S296 by densitometry analysis using NIH ImageJ, normalized to their respective CHK1 levels and compared to the no treatment control group (NT), which was set to 1. (H) Proposed model of CHK1 activation. Following DNA damage and ATR-mediated phosphorylation of CHK1 at S345, CHK1 associates directly or indirectly with KRCC1 and 14-3-3. We posit that this association induces a conformational change in CHK1 to favor autophosphorylation at S296 and enhance kinase activity toward CDC25A.

Article Snippet: The following primary antibodies were used: pCHK1-S345 (2348), pCHK1S296 (2349), H2AX (2577), CHK1 (2360), HA-tag (3724), pH3-S10 (3377) and pan 14-3-3 (8312) from Cell Signaling Technology (Danvers, MA, USA); KRCC1 (16916-1- AP) from Proteintech (Rosemont, IL, USA); CDC25A (sc7389) and CDC7 (sc-56275) from Santa Cruz Biotechnol- ogy (Dallas, TX, USA); pRPA-S33 (A300-246A), AND1 (A301-141A), PSF3 (A304-124A), CHK1 (A300-298A) and pMCM2-S40/41 (A300-788A) from Bethyl Laboratories (Montgomery, TX, USA); DBF4 (ab124707) and pCyclinB1-S126 (ab55184) from Abcam; anti-Halo-tag (G921A) from Promega; RPA32 (MABE285) from EMD Millipore; RAD51 (NB100-148) from Novus Biological; 53BP1 (88439) from Cell Signaling Technology; and - tubulin and -actin from Sigma–Aldrich.

Techniques: Activation Assay, Transfection, Control, Staining, Fluorescence, Microscopy, Western Blot, Immunoprecipitation, Marker, Phospho-proteomics, Activity Assay

Figure 3. Silencing KRCC1 results in delayed S-phase progression and accumulation of cells at the late S phase. Asynchronous HeLa and U2OS cells transfected with control or KRCC1 siRNA and labeled with 20 M EdU for 15 min. DNA synthesis, DNA content and cell cycle distribution were assessed by flow cytometry. (A) Experimental images of the EdU/PI distribution of control and KRCC1 silenced cells. (B) Percentage of cells at the late S– G2 boundary was calculated as a fraction of % EdU-positive cells in the small gate over % total EdU-positive cells. (C) HeLa cells transfected with control or KRCC1 siRNA or treated with CHK1i (AZD7762) or CDC7 inhibitor (CDC7i, TAK-931) were labeled with EdU and subjected to flow cytometry. (D) HeLa cells transfected with control or KRCC1 siRNA or treated with CDC7i (TAK-931) were G1–S synchronized by double thymidine block and released for 12 h. The cells were collected at the indicated time points and analyzed by flow cytometry. (E) Immunoblotting of indicated proteins in control, KRCC1 depleted, CHK1 inhibited or CDC7 inhibited cells. SE and LE blots for pMCM2-S40/41 are shown.

Journal: Nucleic acids research

Article Title: KRCC1, a modulator of the DNA damage response.

doi: 10.1093/nar/gkac890

Figure Lengend Snippet: Figure 3. Silencing KRCC1 results in delayed S-phase progression and accumulation of cells at the late S phase. Asynchronous HeLa and U2OS cells transfected with control or KRCC1 siRNA and labeled with 20 M EdU for 15 min. DNA synthesis, DNA content and cell cycle distribution were assessed by flow cytometry. (A) Experimental images of the EdU/PI distribution of control and KRCC1 silenced cells. (B) Percentage of cells at the late S– G2 boundary was calculated as a fraction of % EdU-positive cells in the small gate over % total EdU-positive cells. (C) HeLa cells transfected with control or KRCC1 siRNA or treated with CHK1i (AZD7762) or CDC7 inhibitor (CDC7i, TAK-931) were labeled with EdU and subjected to flow cytometry. (D) HeLa cells transfected with control or KRCC1 siRNA or treated with CDC7i (TAK-931) were G1–S synchronized by double thymidine block and released for 12 h. The cells were collected at the indicated time points and analyzed by flow cytometry. (E) Immunoblotting of indicated proteins in control, KRCC1 depleted, CHK1 inhibited or CDC7 inhibited cells. SE and LE blots for pMCM2-S40/41 are shown.

Article Snippet: The following primary antibodies were used: pCHK1-S345 (2348), pCHK1S296 (2349), H2AX (2577), CHK1 (2360), HA-tag (3724), pH3-S10 (3377) and pan 14-3-3 (8312) from Cell Signaling Technology (Danvers, MA, USA); KRCC1 (16916-1- AP) from Proteintech (Rosemont, IL, USA); CDC25A (sc7389) and CDC7 (sc-56275) from Santa Cruz Biotechnol- ogy (Dallas, TX, USA); pRPA-S33 (A300-246A), AND1 (A301-141A), PSF3 (A304-124A), CHK1 (A300-298A) and pMCM2-S40/41 (A300-788A) from Bethyl Laboratories (Montgomery, TX, USA); DBF4 (ab124707) and pCyclinB1-S126 (ab55184) from Abcam; anti-Halo-tag (G921A) from Promega; RPA32 (MABE285) from EMD Millipore; RAD51 (NB100-148) from Novus Biological; 53BP1 (88439) from Cell Signaling Technology; and - tubulin and -actin from Sigma–Aldrich.

Techniques: Transfection, Control, Labeling, DNA Synthesis, Flow Cytometry, Blocking Assay, Western Blot

Figure 5. Model of the role of KRCC1 in genome maintenance. Following replication stress and DNA damage, ATR phosphorylates CHK1 at S345. KRCC1 then associates directly or indirectly with CHK1 and 14-3-3 to promote autophosphorylation of CHK1 at S296 and facilitate kinase activity toward CDC25A. CDC25A is then targeted for proteasomal degradation to induce a CHK1-mediated checkpoint. Failure to fully activate CHK1 may result in reduced HRR. We speculate that the KRCC1–CDC7 axis may promote CDC7-mediated events and overall replication integrity, disruption of which leads to replication stress. Overall, replication defects and failure to fully activate checkpoint may result in premature mitotic entry and subsequent apoptosis.

Journal: Nucleic acids research

Article Title: KRCC1, a modulator of the DNA damage response.

doi: 10.1093/nar/gkac890

Figure Lengend Snippet: Figure 5. Model of the role of KRCC1 in genome maintenance. Following replication stress and DNA damage, ATR phosphorylates CHK1 at S345. KRCC1 then associates directly or indirectly with CHK1 and 14-3-3 to promote autophosphorylation of CHK1 at S296 and facilitate kinase activity toward CDC25A. CDC25A is then targeted for proteasomal degradation to induce a CHK1-mediated checkpoint. Failure to fully activate CHK1 may result in reduced HRR. We speculate that the KRCC1–CDC7 axis may promote CDC7-mediated events and overall replication integrity, disruption of which leads to replication stress. Overall, replication defects and failure to fully activate checkpoint may result in premature mitotic entry and subsequent apoptosis.

Article Snippet: The following primary antibodies were used: pCHK1-S345 (2348), pCHK1S296 (2349), H2AX (2577), CHK1 (2360), HA-tag (3724), pH3-S10 (3377) and pan 14-3-3 (8312) from Cell Signaling Technology (Danvers, MA, USA); KRCC1 (16916-1- AP) from Proteintech (Rosemont, IL, USA); CDC25A (sc7389) and CDC7 (sc-56275) from Santa Cruz Biotechnol- ogy (Dallas, TX, USA); pRPA-S33 (A300-246A), AND1 (A301-141A), PSF3 (A304-124A), CHK1 (A300-298A) and pMCM2-S40/41 (A300-788A) from Bethyl Laboratories (Montgomery, TX, USA); DBF4 (ab124707) and pCyclinB1-S126 (ab55184) from Abcam; anti-Halo-tag (G921A) from Promega; RPA32 (MABE285) from EMD Millipore; RAD51 (NB100-148) from Novus Biological; 53BP1 (88439) from Cell Signaling Technology; and - tubulin and -actin from Sigma–Aldrich.

Techniques: Activity Assay, Disruption