Product: Phospho-Chk2 (Thr68) Antibody
Catalog: AF3036
Description: Rabbit polyclonal antibody to Phospho-Chk2 (Thr68)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 62kDa; 61kD(Calculated).
Uniprot: O96017
RRID: AB_2834325

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(86%), Horse(86%), Sheep(86%), Rabbit(100%), Dog(86%)
Clonality:
Polyclonal
Specificity:
Phospho-Chk2 (Thr68) Antibody detects endogenous levels of Chk2 only when phosphorylated at Threonine 68.
RRID:
AB_2834325
Cite Format: Affinity Biosciences Cat# AF3036, RRID:AB_2834325.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

CDS 1; Cds1; Cds1 homolog; Checkpoint kinase 2; Checkpoint like protein CHK2; CHEK 2; Chek2; Chk 2; CHK2 checkpoint homolog (S. pombe); CHK2 checkpoint homolog; CHK2_HUMAN; hCds1; HuCds 1; LFS 2; LFS2; PP1425; RAD 53; RAD53; Rad53 homolog; Serine/threonine protein kinase Chk2; Serine/threonine-protein kinase Chk2;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
O96017 CHK2_HUMAN:

High expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.

Description:
In response to DNA damage and replication blocks, cell cycle progression is halted through the control of critical cell cycle regulators. The protein encoded by this gene is a cell cycle checkpoint regulator and putative tumor suppressor.
Sequence:
MSRESDVEAQQSHGSSACSQPHGSVTQSQGSSSQSQGISSSSTSTMPNSSQSSHSSSGTLSSLETVSTQELYSIPEDQEPEDQEPEEPTPAPWARLWALQDGFANLECVNDNYWFGRDKSCEYCFDEPLLKRTDKYRTYSKKHFRIFREVGPKNSYIAYIEDHSGNGTFVNTELVGKGKRRPLNNNSEIALSLSRNKVFVFFDLTVDDQSVYPKALRDEYIMSKTLGSGACGEVKLAFERKTCKKVAIKIISKRKFAIGSAREADPALNVETEIEILKKLNHPCIIKIKNFFDAEDYYIVLELMEGGELFDKVVGNKRLKEATCKLYFYQMLLAVQYLHENGIIHRDLKPENVLLSSQEEDCLIKITDFGHSKILGETSLMRTLCGTPTYLAPEVLVSVGTAGYNRAVDCWSLGVILFICLSGYPPFSEHRTQVSLKDQITSGKYNFIPEVWAEVSEKALDLVKKLLVVDPKARFTTEEALRHPWLQDEDMKRKFQDLLSEENESTALPQVLAQPSTSRKRPREGEAEGAETTKRPAVCAAVL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Rabbit
100
Pig
100
Horse
86
Bovine
86
Sheep
86
Dog
86
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O96017 As Substrate

Site PTM Type Enzyme
S12 Phosphorylation
S15 Phosphorylation
S19 Phosphorylation O96017 (CHEK2) , Q13315 (ATM)
S24 Phosphorylation
T26 Phosphorylation P53350 (PLK1) , Q13315 (ATM)
S28 Phosphorylation Q13315 (ATM)
S33 Phosphorylation Q13315 (ATM) , O96017 (CHEK2)
S35 Phosphorylation O96017 (CHEK2) , Q13315 (ATM)
S39 Phosphorylation
S40 Phosphorylation
S41 Phosphorylation
S42 Phosphorylation
T43 Phosphorylation
S44 Phosphorylation
T45 Phosphorylation
S50 Phosphorylation Q13315 (ATM)
S52 Phosphorylation
S55 Phosphorylation
S62 Phosphorylation Q9H4B4 (PLK3)
T65 Phosphorylation
S67 Phosphorylation
T68 Phosphorylation P33981 (TTK) , P78527 (PRKDC) , Q13535 (ATR) , P53350 (PLK1) , Q9NYL2 (MAP3K20) , Q13315 (ATM) , O96017 (CHEK2) , Q683Z8 (CHK2)
S73 Phosphorylation Q9H4B4 (PLK3)
K119 Ubiquitination
S120 Phosphorylation O96017 (CHEK2)
K131 Ubiquitination
S140 Phosphorylation O96017 (CHEK2)
S164 Phosphorylation P53350 (PLK1)
T205 Phosphorylation P53350 (PLK1)
S210 Phosphorylation P53350 (PLK1)
K224 Ubiquitination
T225 Phosphorylation O96017 (CHEK2)
K235 Acetylation
S260 Phosphorylation O96017 (CHEK2)
K279 Ubiquitination
K287 Ubiquitination
S357 Phosphorylation Q13315 (ATM)
S372 Phosphorylation
K373 Ubiquitination
T378 Phosphorylation
S379 Phosphorylation O96017 (CHEK2)
T383 Phosphorylation P78527 (PRKDC) , O96017 (CHEK2) , Q683Z8 (CHK2)
T387 Phosphorylation Q683Z8 (CHK2) , P78527 (PRKDC) , O96017 (CHEK2)
T389 Phosphorylation
Y390 Phosphorylation
T432 Phosphorylation O96017 (CHEK2)
S435 Phosphorylation O96017 (CHEK2)
K437 Ubiquitination
K444 Ubiquitination
S456 Phosphorylation
K458 Ubiquitination
K472 Ubiquitination
K494 Ubiquitination
S516 Phosphorylation O96017 (CHEK2) , Q683Z8 (CHK2)
T517 Phosphorylation
C539 S-Nitrosylation

PTMs - O96017 As Enzyme

Substrate Site Source
O00716 (E2F3) S124 Uniprot
O15151 (MDM4) S342 Uniprot
O15151-1 (MDM4) S367 Uniprot
O43255 (SIAH2) T26 Uniprot
O43255 (SIAH2) S28 Uniprot
O43255 (SIAH2) S68 Uniprot
O43255 (SIAH2) T119 Uniprot
O60673 (REV3L) S1075 Uniprot
O96017 (CHEK2) S19 Uniprot
O96017 (CHEK2) S33 Uniprot
O96017 (CHEK2) S35 Uniprot
O96017-12 (CHEK2) T68 Uniprot
O96017 (CHEK2) S120 Uniprot
O96017 (CHEK2) S140 Uniprot
O96017 (CHEK2) T225 Uniprot
O96017 (CHEK2) S260 Uniprot
O96017-12 (CHEK2) T354 Uniprot
O96017-12 (CHEK2) T358 Uniprot
O96017 (CHEK2) S379 Uniprot
O96017 (CHEK2) T383 Uniprot
O96017 (CHEK2) T387 Uniprot
O96017-9 (CHEK2) T426 Uniprot
O96017-9 (CHEK2) T430 Uniprot
O96017 (CHEK2) T432 Uniprot
O96017 (CHEK2) S435 Uniprot
O96017-12 (CHEK2) S487 Uniprot
O96017-1 (CHEK2) S516 Uniprot
O96017-9 (CHEK2) S559 Uniprot
P04637 (TP53) S15 Uniprot
P04637 (TP53) T18 Uniprot
P04637 (TP53) S20 Uniprot
P04637 (TP53) S37 Uniprot
P04637 (TP53) S366 Uniprot
P04637 (TP53) S378 Uniprot
P06400 (RB1) S612 Uniprot
P10636-8 (MAPT) S262 Uniprot
P10636 (MAPT) S579 Uniprot
P16403 (HIST1H1C) S36 Uniprot
P16403 (HIST1H1C) S55 Uniprot
P16403 (HIST1H1C) S89 Uniprot
P16403 (HIST1H1C) T92 Uniprot
P16403 (HIST1H1C) T167 Uniprot
P18887 (XRCC1) S184 Uniprot
P18887 (XRCC1) S210 Uniprot
P18887 (XRCC1) T284 Uniprot
P21127 (CDK11B) S752 Uniprot
P23560 (BDNF) T62 Uniprot
P29590-3 (PML) S117 Uniprot
P30304 (CDC25A) S124 Uniprot
P30304 (CDC25A) S178 Uniprot
P30304-2 (CDC25A) S239 Uniprot
P30304-2 (CDC25A) S253 Uniprot
P30304 (CDC25A) S279 Uniprot
P30304 (CDC25A) S293 Uniprot
P30307 (CDC25C) S216 Uniprot
P33981 (TTK) T288 Uniprot
P38398-8 (BRCA1) S941 Uniprot
P38398-7 (BRCA1) S988 Uniprot
P40337 (VHL) S111 Uniprot
P54132 (BLM) T182 Uniprot
P61289 (PSME3) S247 Uniprot
Q01094 (E2F1) S364 Uniprot
Q08050-2 (FOXM1) S361 Uniprot
Q08050 (FOXM1) S376 Uniprot
Q13263 (TRIM28) S473 Uniprot
Q13972 (RASGRF1) S287 Uniprot
Q15717 (ELAVL1) S88 Uniprot
Q15717 (ELAVL1) S100 Uniprot
Q15717 (ELAVL1) T118 Uniprot
Q9NRM7 (LATS2) S172 Uniprot
Q9NRM7 (LATS2) S380 Uniprot
Q9NRM7 (LATS2) S408 Uniprot
Q9NRM7 (LATS2) S446 Uniprot
Q9NY61 (AATF) S141 Uniprot
Q9NY61 (AATF) S143 Uniprot
Q9NY61 (AATF) S477 Uniprot
Q9NY61 (AATF) S510 Uniprot
Q9UQ88 (CDK11A) S740 Uniprot

Research Backgrounds

Function:

Serine/threonine-protein kinase which is required for checkpoint-mediated cell cycle arrest, activation of DNA repair and apoptosis in response to the presence of DNA double-strand breaks. May also negatively regulate cell cycle progression during unperturbed cell cycles. Following activation, phosphorylates numerous effectors preferentially at the consensus sequence [L-X-R-X-X-S/T]. Regulates cell cycle checkpoint arrest through phosphorylation of CDC25A, CDC25B and CDC25C, inhibiting their activity. Inhibition of CDC25 phosphatase activity leads to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. May also phosphorylate NEK6 which is involved in G2/M cell cycle arrest. Regulates DNA repair through phosphorylation of BRCA2, enhancing the association of RAD51 with chromatin which promotes DNA repair by homologous recombination. Also stimulates the transcription of genes involved in DNA repair (including BRCA2) through the phosphorylation and activation of the transcription factor FOXM1. Regulates apoptosis through the phosphorylation of p53/TP53, MDM4 and PML. Phosphorylation of p53/TP53 at 'Ser-20' by CHEK2 may alleviate inhibition by MDM2, leading to accumulation of active p53/TP53. Phosphorylation of MDM4 may also reduce degradation of p53/TP53. Also controls the transcription of pro-apoptotic genes through phosphorylation of the transcription factor E2F1. Tumor suppressor, it may also have a DNA damage-independent function in mitotic spindle assembly by phosphorylating BRCA1. Its absence may be a cause of the chromosomal instability observed in some cancer cells. Promotes the CCAR2-SIRT1 association and is required for CCAR2-mediated SIRT1 inhibition.

PTMs:

Phosphorylated. Phosphorylated at Ser-73 by PLK3 in response to DNA damage, promoting phosphorylation at Thr-68 by ATM and the G2/M transition checkpoint. Phosphorylation at Thr-68 induces homodimerization. Autophosphorylates at Thr-383 and Thr-387 in the T-loop/activation segment upon dimerization to become fully active and phosphorylate its substrates like for instance CDC25C. DNA damage-induced autophosphorylation at Ser-379 induces CUL1-mediated ubiquitination and regulates the pro-apoptotic function. Phosphorylation at Ser-456 also regulates ubiquitination. Phosphorylated by PLK4.

Ubiquitinated. CUL1-mediated ubiquitination regulates the pro-apoptotic function. Ubiquitination may also regulate protein stability. Ubiquitinated by RNF8 via 'Lys-48'-linked ubiquitination.

Subcellular Location:

Nucleus.
Note: Isoform 10 is present throughout the cell.

Nucleus.

Nucleus.

Nucleus.

Nucleus.

Nucleus>PML body. Nucleus>Nucleoplasm.
Note: Recruited into PML bodies together with TP53.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

High expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.

Subunit Structure:

Homodimer. Homodimerization is part of the activation process but the dimer may dissociate following activation. Interacts with PML. Interacts with TP53. Interacts with RB1; phosphorylates RB1. Interacts with BRCA1. Interacts (phosphorylated at Thr-68) with MDC1; requires ATM-mediated phosphorylation of CHEK2. Interacts with TP53BP1; modulates CHEK2 phosphorylation at Thr-68 in response to ionizing radiation. Interacts with CDC25A; phosphorylates CDC25A and mediates its degradation in response to ionizing radiation. Interacts with CUL1; mediates CHEK2 ubiquitination and regulation. Interacts with CDKN2AIP. Interacts (via protein kinase domain) with CCAR2 (via N-terminus). Interacts with SIRT1.

Family&Domains:

Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHK2 subfamily.

Research Fields

· Cellular Processes > Cell growth and death > Cell cycle.   (View pathway)

· Cellular Processes > Cell growth and death > p53 signaling pathway.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

References

1). The activated ATM/p53 pathway promotes autophagy in response to oxidative stress-mediated DNA damage induced by Microcystin-LR in male germ cells. Ecotoxicology and Environmental Safety (PubMed: 34715501) [IF=6.8]

Application: WB    Species: Mouse    Sample: GC-1 cells

Fig. 4. Changes of ATM and its downstream proteins in GC-1 cells and mouse testis. Expression and analysis of the related proteins in mouse testis (A) and in GC-1 cells (B). *p < 0.05 vs. the control group; #p < 0.05 vs. the corresponding MC-LR exposure group. All data were expressed as  ± SD (n = 3).

2). Role of DNA damage in the progress of chronic tubule‑interstitial injury. Molecular Medicine Reports (PubMed: 32626982) [IF=3.4]

Application: WB    Species: human    Sample: HK-2 cells

Figure 4.| - p21 knockdown reduces the DNA damage response in HK-2 cells. (A) Protein expression levels of p-ATM, ATM, p-Chk2, Chk2, rH2AX, p-p53 and p53 were analyzed using western blotting in cells transfected with shp21 or shCon with or without AA-induced injury.

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