Antibodies for Cell Cycle Checkpoint Kinases CHK1, CHK2 - FocusOn 033

Acris Antibodies Panel to CHK1 and CHK2

Acris Antibodies offers monoclonal and polyclonal antibodies to CHK1 and CHK2, which were raised against recombinant or synthetic CHK, and to the phosphorylated forms using phosphopeptides for immunization (see Product list).

Introduction

Cell cycle events are regulated by the sequential activation and deactivation of cyclin dependent kinases (Cdks) and by proteolysis of cyclins. Serine/threonine-protein kinases CHK 1 and CHK 2 are involved in these processes as regulators of Cdks.

CHK 1 and CHK 2 both function as essential components in the G2 DNA damage checkpoint by phosphorylating CDC25C in response to DNA damage. CDC25A, CDC25B and CDC25C protein tyrosine phosphatases function as mitotic activators by dephosphorylating CDC2 p34 on regulatory tyrosine residues. Phosphorylation of CDC25C by CHK 1 and 2 inhibits its phosphatase activity, thereby blocking mitosis.

CHK1

It has been shown that CHK1 can phosphorylate Wee 1 in vitro, providing evidence that the hyperphosphorylated form of Wee 1, seen in cells delayed by CHK1 over-expression, is due to phosphorylation by CHK1.

CHK2

The p53 tumor-suppressor gene integrates numerous signals that control cell life and death. Several novel molecules involved in p53 signalling, including CHK2, p53R2, p53AIP1, Noxa, PIDD, and PID/MTA2, have been discovered. The checkpoint kinase CHK2 is the mammalian homologue of yeast Cds1/Rad53. In response to DNA damage, the checkpoint kinase ATM phosphorylates and activates CHK2, which in turn directly phosphorylates and activates p53. CHK2 serves an ATM downstream effector to mediate activation of p53. CHK2 also phosphorylates and activates BRCA1, the product of a tumor suppressor gene that is mutated in breast and ovarian cancer.

Fig. 1: Formalin-fixed and paraffin-embedded (FFPE) human cancer tissue reacted with the CHK1 antibody Cat.-No. AP14043PU-N; peroxidase-conjugated secondary antibody followed by DAB staining
Fig. 2: Cat.-No. AP12591PU-N is used in WB to detect Phospho-CHK1 (pSer280) in A2058, mouse brain, Ramos, Jurkat, HL60, and Hela tissue lysates (from left to right)
Fig. 3: CHK1 antibody (pSer317) Cat.-No. AP19048PU-N staining of paraformaldehyde-fixed HeLa by immunofluorescence at 1:200 dilution
Fig. 4: CHK1 antibody Cat.-No. AP05017PU-N staining of cell lines (RKO, HCT116,COS) at 1:500 dilution
Fig. 5: Human testis (FFPE) stained with CHK2 antibody Cat.-No. AP07293PU-N at 5 µg/l; biotinylated secondary antibody, alkaline phosphatase-streptavidin and chromogen
Fig. 6: WB using CHK2 (pThr68) antibody Cat.-No. R1536 shows detection of a predominant band at ~60 kDa corresponding to phosphorylated CHK2 (arrowhead) in MCF-7 whole cell lysates after treatment with doxorubicin. CHK2 phosphorylation was induced using increasing concentrations of the DNA damaging agent doxorubicin as indicated for 24 h prior to lysate production. Personal communication, Xiao HeYang, University of Oklahoma Health Sciences Center

Key references

 
Allinen, M et al: Mutation analysis of the CHK2 gene in families with hereditary breast cancer.
Br J Cancer, 2001; 85 (2): 209-212.
 
Bell, DW et al: Heterozygous germ line hCHK2 mutations in Li-Fraumeni syndrome.
Science, 1999; 286 (5449): 2528-2531.
 
Dong, X et. al: Mutations in CHEK2 associated with prostate cancer risk.
Am J Hum Genet, 2003; 72 (2): 270-280.
 
Gonzales S et al: p73 alpha regulation by Chk1 in response to DNA damage.
Mol Cell Biol, Nov 2003; 23(22): 8161-71.
 
Miller, CW et al: Mutations of the CHK2 gene are found in some osteosarcomas, but are rare in breast, lung, and ovarian tumors.
Genes Chromosomes Cancer, 2002; 33 (1): 17-21.
 
Storgaard Sørensen, C et al: The cell cycle checkpoint kinase Chk1 is required for mammalian homologous recombination repair.
Nature Cell Biology, 2005; 7: 195-201.

Further readings

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

CHK1 antibody

AP32963PU-N (2µg/ml) staining of Rat Thymus lysate (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence. Goat Aff - Purified Bov, Hu, Ms, Rt WB
0.1 mg / €340.00
  Acris Antibodies GmbH

CHK1 antibody

Western blot analysis using CHK1 antibody Cat.-No AM06099PU-N against A431 (Lane 1), Hela (Lane 2), NIH/3T3 (Lane 3) and K562 (Lane 4) cell lysate. Mouse IgG1 2G1D5 Purified Hu, Ms E, WB
0.1 ml / €330.00
  Acris Antibodies GmbH

CHK1 antibody

Figure 1. Western blot analysis of anti-CHK1 Mab (Cat. #AM11058SU-N) to detect CHK1 in NIH/3T3 cell lysate (Lane 1) and K562 cell lysate (Lane 2). Mouse 2G1D5 Purified Hu, Ms WB
0.4 ml / €370.00
  Acris Antibodies GmbH

CHK1 antibody

Figure 1. AM20031AF-N CHK1 antibody staining of Paraffin Embedded Human Tonsil Section (germinal center). Mouse IgG2b DCS-310 Azide Free Hu IP, P, WB
0.1 mg / €340.00
  Acris Antibodies GmbH

CHK1 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Chk1 antibody AP02645PU. Rabbit Aff - Purified P, WB
0.1 ml / €270.00
  Acris Antibodies GmbH

CHK1 antibody

AP05017PU-N CHK1 antibody staining of cell lines at 1/500 dilution. Sheep IgG Purified Hu, Mky, Ms, Rt IP, WB
0.25 mg / €360.00
  Acris Antibodies GmbH

CHK1 pSer280 antibody

Thymus: Formalin-Fixed Paraffin-Embedded (FFPE) Rabbit IgG Aff - Purified Hu P, WB
50 µg / €470.00
  Acris Antibodies GmbH

CHK1 (312-327) antibody

Western blot using Affinity Purified anti-Yeast CHK1 antibody shows detection of a bands corresponding to CHK1 in Saccharomyces cerevisiae lysates. Two strains of S.cerevisiae were tested. Lane 1 shows a predominant band at ~60 kDa. Lane 2 shows a predominant band at ~38 kDa. Specific band staining is blocked when antibody is preincubated for 45 min at room temperature with 50 µg of peptide immunogen (lanes 3 and 4 respectively). Lysates were separated by 4-20% SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed for 2 h at room temperature with the primary antibody diluted to 1:750 in blocking buffer diluted 1:5 in PBS. The membrane was washed and reacted with a 1:10,000 dilution of IRDye(TM)800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). IRDye(TM)800 fluorescence image was captured using the Odyssey(R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results. Rabbit IgG Aff - Purified Ye E, WB
0.1 mg / €440.00
  Acris Antibodies GmbH

CHK1 pSer280 antibody

Western blot analysis of extracts from A2780 cells treated with calf intestinal phosphatase (CIP using Chk1 (phospho-Ser280) antibody (Cat.-No AP09458PU) Rabbit Aff - Purified Hu WB
0.1 ml / €320.00
  Acris Antibodies GmbH

CHK1 pSer280 antibody

Western blot analysis of extracts from A2780 cells treated with calf intestinal phosphatase (CIP using Chk1 (phospho-Ser280) antibody (Cat.-No AP09458PU) Rabbit Aff - Purified Hu WB
50 µl / €240.00
  Acris Antibodies GmbH

CHK1 antibody

Western blot analysis of anti-CHK1 Antibody (S280) (Cat.#AP12111PU-N) in Hela cell line lysates (35ug/lane). CHK1 (arrow) was detected using the purified Pab. Rabbit Ig Aff - Purified Hu WB
0.4 ml / €370.00
  Acris Antibodies GmbH

CHK1 pSer280 antibody

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Aff - Purified Hu, Ms P, WB

Regular Price: 0.4 ml / €370.00

Special Price: 0.4 ml / €285.00

  Acris Antibodies GmbH

CHK1 pSer317 antibody

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Aff - Purified Hu P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

CHK1 antibody

Detection of human Chk1 in whole cell lysates of HeLa cells by immunoprecipitation and Western blotting using Goat anti Human Chk1(SP2056P). Immunprecipitated material was detected in Western blotting with a monoclonal anti Chk1 antibody Goat Aff - Purified Hu IP, WB
50 µg / €330.00
  Acris Antibodies GmbH

CHK2 antibody

Figure 1. AM00536AF-N CHK2 antibody Immunohistochemical staining of Human Intestine Paraffin Embedded Section Mouse IgG2a DCS-270 Azide Free Hu IP, P, WB
0.1 mg / €320.00
  Acris Antibodies GmbH

CHK2 antibody

AM06003PU-N CHEK2 antibody staining of Formalin-Fixed,  Paraffin-Embedded Human Spleen at 10 µg/ml followed by Biotin anti-Mouse IgG secondary antibody, Alkaline Phosphatase-streptavidin and Chromogen. Mouse IgG1 73C175.1.1 Purified Hu P, WB
50 µg / €470.00
  Acris Antibodies GmbH

CHK2 antibody

Immunohistochemical analysis of paraffin-embedded human lung carcinoma (A), liver carcinoma (B), breast carcinoma (C) and kiney carcinoma (D), showing nuclear localization with DAB staining using CHK2 antibody Cat.-No AM06242SU-N Mouse IgG2b 1C12B8 Ascites Hu E, ICC/IF, P, WB
0.1 ml / €460.00
  Acris Antibodies GmbH

CHK2 antibody

Figure 1. AM20032AF-N CHK2 antibody Immunohistochemical staining of Human Intestine Paraffin Embedded Section. Mouse IgG2a DCS-273 Azide Free Hu IP, P, WB
0.1 mg / €320.00
  Acris Antibodies GmbH

CHK2 pSer516 antibody

Figure 1. Western blot analysis of extracts from HeLa cells using Chk2 (phospho-Ser516) antibody (AP02432PU). Rabbit Aff - Purified Hu WB
0.1 ml / €320.00
  Acris Antibodies GmbH

CHK2 pSer516 antibody

Figure 1. Western blot analysis of extracts from HeLa cells using Chk2 (phospho-Ser516) antibody (AP02432PU). Rabbit Aff - Purified Hu WB
50 µl / €240.00
  Acris Antibodies GmbH

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