Phospho-ATR (Ser428) Antibody - #DF7512

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Product: Phospho-ATR (Ser428) Antibody
Catalog: DF7512
Description: Rabbit polyclonal antibody to Phospho-ATR (Ser428)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Horse, Rabbit, Dog
Mol.Wt.: 300 kDa; 301kD(Calculated).
Uniprot: Q13535
RRID: AB_2841011

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Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:50-1:200
*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(88%), Horse(100%), Rabbit(100%), Dog(83%)
Clonality:
Polyclonal
Specificity:
Phospho-ATR (Ser428) Antibody detects endogenous levels of ATR only when phosphorylated at S428.
RRID:
AB_2841011
Cite Format: Affinity Biosciences Cat# DF7512, RRID:AB_2841011.
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

Ataxia telangiectasia and Rad3 related; Ataxia telangiectasia and Rad3-related protein; ATR; ATR_HUMAN; FCTCS; FRAP Related Protein 1; FRAP-related protein 1; FRP1; MEC1; MEC1 mitosis entry checkpoint 1 homolog; Protein kinase ATR; RAC3; Rad3 related protein; SCKL; SCKL1; Serine/threonine protein kinase ATR; Serine/threonine-protein kinase ATR;

Immunogens

Immunogen:
Uniprot:

Q13535(ATR_HUMAN) >>Visit HPA database.

Gene(ID):
ATR(545)
Expression:
Q13535 ATR_HUMAN:

Ubiquitous, with highest expression in testis. Isoform 2 is found in pancreas, placenta and liver but not in heart, testis and ovary.

Sequence:
MGEHGLELASMIPALRELGSATPEEYNTVVQKPRQILCQFIDRILTDVNVVAVELVKKTDSQPTSVMLLDFIQHIMKSSPLMFVNVSGSHEAKGSCIEFSNWIITRLLRIAATPSCHLLHKKICEVICSLLFLFKSKSPAIFGVLTKELLQLFEDLVYLHRRNVMGHAVEWPVVMSRFLSQLDEHMGYLQSAPLQLMSMQNLEFIEVTLLMVLTRIIAIVFFRRQELLLWQIGCVLLEYGSPKIKSLAISFLTELFQLGGLPAQPASTFFSSFLELLKHLVEMDTDQLKLYEEPLSKLIKTLFPFEAEAYRNIEPVYLNMLLEKLCVMFEDGVLMRLKSDLLKAALCHLLQYFLKFVPAGYESALQVRKVYVRNICKALLDVLGIEVDAEYLLGPLYAALKMESMEIIEEIQCQTQQENLSSNSDGISPKRRRLSSSLNPSKRAPKQTEEIKHVDMNQKSILWSALKQKAESLQISLEYSGLKNPVIEMLEGIAVVLQLTALCTVHCSHQNMNCRTFKDCQHKSKKKPSVVITWMSLDFYTKVLKSCRSLLESVQKLDLEATIDKVVKIYDALIYMQVNSSFEDHILEDLCGMLSLPWIYSHSDDGCLKLTTFAANLLTLSCRISDSYSPQAQSRCVFLLTLFPRRIFLEWRTAVYNWALQSSHEVIRASCVSGFFILLQQQNSCNRVPKILIDKVKDDSDIVKKEFASILGQLVCTLHGMFYLTSSLTEPFSEHGHVDLFCRNLKATSQHECSSSQLKASVCKPFLFLLKKKIPSPVKLAFIDNLHHLCKHLDFREDETDVKAVLGTLLNLMEDPDKDVRVAFSGNIKHILESLDSEDGFIKELFVLRMKEAYTHAQISRNNELKDTLILTTGDIGRAAKGDLVPFALLHLLHCLLSKSASVSGAAYTEIRALVAAKSVKLQSFFSQYKKPICQFLVESLHSSQMTALPNTPCQNADVRKQDVAHQREMALNTLSEIANVFDFPDLNRFLTRTLQVLLPDLAAKASPAASALIRTLGKQLNVNRREILINNFKYIFSHLVCSCSKDELERALHYLKNETEIELGSLLRQDFQGLHNELLLRIGEHYQQVFNGLSILASFASSDDPYQGPRDIISPELMADYLQPKLLGILAFFNMQLLSSSVGIEDKKMALNSLMSLMKLMGPKHVSSVRVKMMTTLRTGLRFKDDFPELCCRAWDCFVRCLDHACLGSLLSHVIVALLPLIHIQPKETAAIFHYLIIENRDAVQDFLHEIYFLPDHPELKKIKAVLQEYRKETSESTDLQTTLQLSMKAIQHENVDVRIHALTSLKETLYKNQEKLIKYATDSETVEPIISQLVTVLLKGCQDANSQARLLCGECLGELGAIDPGRLDFSTTETQGKDFTFVTGVEDSSFAYGLLMELTRAYLAYADNSRAQDSAAYAIQELLSIYDCREMETNGPGHQLWRRFPEHVREILEPHLNTRYKSSQKSTDWSGVKKPIYLSKLGSNFAEWSASWAGYLITKVRHDLASKIFTCCSIMMKHDFKVTIYLLPHILVYVLLGCNQEDQQEVYAEIMAVLKHDDQHTINTQDIASDLCQLSTQTVFSMLDHLTQWARHKFQALKAEKCPHSKSNRNKVDSMVSTVDYEDYQSVTRFLDLIPQDTLAVASFRSKAYTRAVMHFESFITEKKQNIQEHLGFLQKLYAAMHEPDGVAGVSAIRKAEPSLKEQILEHESLGLLRDATACYDRAIQLEPDQIIHYHGVVKSMLGLGQLSTVITQVNGVHANRSEWTDELNTYRVEAAWKLSQWDLVENYLAADGKSTTWSVRLGQLLLSAKKRDITAFYDSLKLVRAEQIVPLSAASFERGSYQRGYEYIVRLHMLCELEHSIKPLFQHSPGDSSQEDSLNWVARLEMTQNSYRAKEPILALRRALLSLNKRPDYNEMVGECWLQSARVARKAGHHQTAYNALLNAGESRLAELYVERAKWLWSKGDVHQALIVLQKGVELCFPENETPPEGKNMLIHGRAMLLVGRFMEETANFESNAIMKKYKDVTACLPEWEDGHFYLAKYYDKLMPMVTDNKMEKQGDLIRYIVLHFGRSLQYGNQFIYQSMPRMLTLWLDYGTKAYEWEKAGRSDRVQMRNDLGKINKVITEHTNYLAPYQFLTAFSQLISRICHSHDEVFVVLMEIIAKVFLAYPQQAMWMMTAVSKSSYPMRVNRCKEILNKAIHMKKSLEKFVGDATRLTDKLLELCNKPVDGSSSTLSMSTHFKMLKKLVEEATFSEILIPLQSVMIPTLPSILGTHANHASHEPFPGHWAYIAGFDDMVEILASLQKPKKISLKGSDGKFYIMMCKPKDDLRKDCRLMEFNSLINKCLRKDAESRRRELHIRTYAVIPLNDECGIIEWVNNTAGLRPILTKLYKEKGVYMTGKELRQCMLPKSAALSEKLKVFREFLLPRHPPIFHEWFLRTFPDPTSWYSSRSAYCRSTAVMSMVGYILGLGDRHGENILFDSLTGECVHVDFNCLFNKGETFEVPEIVPFRLTHNMVNGMGPMGTEGLFRRACEVTMRLMRDQREPLMSVLKTFLHDPLVEWSKPVKGHSKAPLNETGEVVNEKAKTHVLDIEQRLQGVIKTRNRVTGLPLSIEGHVHYLIQEATDENLLCQMYLGWTPYM

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
Horse
100
Pig
88
Dog
83
Bovine
75
Sheep
67
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13535 As Substrate

Site PTM Type Enzyme
K32 Ubiquitination
S61 Phosphorylation
R177 Methylation
Y291 Phosphorylation P00519 (ABL1)
S296 Phosphorylation
K297 Ubiquitination
K300 Ubiquitination
Y310 Phosphorylation P00519 (ABL1)
S428 Phosphorylation P06493 (CDK1)
S435 Phosphorylation P17612 (PRKACA)
S436 Phosphorylation
S437 Phosphorylation
S441 Phosphorylation
K442 Ubiquitination
K459 Ubiquitination
S464 Phosphorylation
T533 Phosphorylation
K695 Ubiquitination
K746 Methylation
K746 Ubiquitination
K759 Methylation
K764 Ubiquitination
K791 Ubiquitination
K818 Ubiquitination
K851 Ubiquitination
K866 Ubiquitination
K918 Ubiquitination
S919 Phosphorylation
K921 Ubiquitination
T952 Phosphorylation
K1005 Ubiquitination
S1007 Phosphorylation
K1019 Ubiquitination
Y1122 Phosphorylation
S1140 Phosphorylation
K1165 Ubiquitination
K1173 Acetylation
K1173 Ubiquitination
K1185 Ubiquitination
T1275 Phosphorylation
K1308 Ubiquitination
S1333 Phosphorylation Q13535 (ATR)
K1467 Ubiquitination
K1475 Ubiquitination
K1476 Ubiquitination
T1566 Phosphorylation
T1578 Phosphorylation
T1589 Phosphorylation
K1600 Ubiquitination
K1613 Ubiquitination
S1645 Phosphorylation
K1666 Ubiquitination
K1697 Ubiquitination
K1703 Ubiquitination
K1824 Ubiquitination
S1871 Phosphorylation
S1876 Phosphorylation
K1897 Ubiquitination
K1912 Ubiquitination
K1933 Ubiquitination
T1989 Phosphorylation Q13535 (ATR)
K1994 Ubiquitination
K2026 Ubiquitination
K2048 Ubiquitination
K2106 Ubiquitination
K2121 Ubiquitination
S2186 Phosphorylation
Y2187 Phosphorylation
K2200 Ubiquitination
K2210 Ubiquitination
K2221 Ubiquitination
K2228 Ubiquitination
S2313 Phosphorylation
K2347 Ubiquitination
K2397 Ubiquitination
K2404 Ubiquitination
K2420 Ubiquitination
K2567 Ubiquitination
K2574 Ubiquitination
K2587 Ubiquitination
K2589 Ubiquitination
K2604 Ubiquitination

PTMs - Q13535 As Enzyme

Substrate Site Source
O00203 (AP3B1) S1092 Uniprot
O14757 (CHEK1) S296 Uniprot
O14757-1 (CHEK1) S317 Uniprot
O14757 (CHEK1) S345 Uniprot
O15360 (FANCA) S1449 Uniprot
O43542 (XRCC3) S225 Uniprot
O60934 (NBN) S343 Uniprot
O60934 (NBN) S615 Uniprot
O75717 (WDHD1) T826 Uniprot
O75943-3 (RAD17) S470 Uniprot
O75943-3 (RAD17) S480 Uniprot
O75943-4 (RAD17) S549 Uniprot
O75943-4 (RAD17) S559 Uniprot
O75943-2 (RAD17) S635 Uniprot
O75943-2 (RAD17) S645 Uniprot
O75943 (RAD17) S646 Uniprot
O75943 (RAD17) S656 Uniprot
O94880 (PHF14) S294 Uniprot
O96017-9 (CHEK2) S19 Uniprot
O96017-12 (CHEK2) T26 Uniprot
O96017-9 (CHEK2) S28 Uniprot
O96017-9 (CHEK2) S33 Uniprot
O96017-12 (CHEK2) S35 Uniprot
O96017-12 (CHEK2) S50 Uniprot
O96017 (CHEK2) T68 Uniprot
P04637 (TP53) S15 Uniprot
P04637-1 (TP53) T18 Uniprot
P04637-1 (TP53) S20 Uniprot
P04637-1 (TP53) S37 Uniprot
P06748 (NPM1) S125 Uniprot
P08047 (SP1) S101 Uniprot
P15927 (RPA2) T21 Uniprot
P15927 (RPA2) S23 Uniprot
P15927 (RPA2) S29 Uniprot
P15927 (RPA2) S33 Uniprot
P15927 (RPA2) S52 Uniprot
P15927 (RPA2) S72 Uniprot
P15927 (RPA2) S174 Uniprot
P16220 (CREB1) S121 Uniprot
P18583 (SON) S1216 Uniprot
P23025 (XPA) S173 Uniprot
P23025 (XPA) S196 Uniprot
P25205 (MCM3) S535 Uniprot
P27707 (DCK) S74 Uniprot
P28340 (POLD1) S717 Uniprot
P30307 (CDC25C) S216 Uniprot
P35568 (IRS1) S270 Uniprot
P38398-6 (BRCA1) T291 Uniprot
P38398-6 (BRCA1) S320 Uniprot
P38398-3 (BRCA1) S353 Uniprot
P38398-8 (BRCA1) S1096 Uniprot
P38398-7 (BRCA1) S1143 Uniprot
P38398-8 (BRCA1) S1233 Uniprot
P38398-7 (BRCA1) S1280 Uniprot
P38398 (BRCA1) S1298 Uniprot
P38398-8 (BRCA1) T1347 Uniprot
P38398-8 (BRCA1) S1376 Uniprot
P38398 (BRCA1) S1387 Uniprot
P38398 (BRCA1) T1394 Uniprot
P38398-8 (BRCA1) S1410 Uniprot
P38398 (BRCA1) S1423 Uniprot
P38398-1 (BRCA1) S1457 Uniprot
P38398 (BRCA1) S1466 Uniprot
P38398 (BRCA1) S1524 Uniprot
P39880 (CUX1) S1357 Uniprot
P40692 (MLH1) S406 Uniprot
P42771 (CDKN2A) S7 Uniprot
P42771 (CDKN2A) S8 Uniprot
P42771 (CDKN2A) S140 Uniprot
P42771 (CDKN2A) S152 Uniprot
P49006 (MARCKSL1) S71 Uniprot
P49736 (MCM2) S108 Uniprot
P49917 (LIG4) S199 Uniprot
P49959 (MRE11) S264 Uniprot
P49959 (MRE11) S382 Uniprot
P49959 (MRE11) S531 Uniprot
P51587 (BRCA2) S2156 Uniprot
P54132 (BLM) T99 Uniprot
P54132 (BLM) T122 Uniprot
P55072 (VCP) S784 Uniprot
P55957 (BID) S78 Uniprot
P78527 (PRKDC) T2609 Uniprot
P78527 (PRKDC) S2612 Uniprot
P78527 (PRKDC) T2647 Uniprot
Q00987 (MDM2) S407 Uniprot
Q01094 (E2F1) S31 Uniprot
Q01844 (EWSR1) T79 Uniprot
Q03164 (KMT2A) S516 Uniprot
Q06190 (PPP2R3A) S61 Uniprot
Q10567 (AP1B1) S936 Uniprot
Q13098 (GPS1) S474 Uniprot
Q13315 (ATM) S440 Uniprot
Q13315 (ATM) S1981 Uniprot
Q13315 (ATM) S2761 Uniprot
Q13535 (ATR) S1333 Uniprot
Q13535 (ATR) T1989 Uniprot
Q13541 (EIF4EBP1) S94 Uniprot
Q14191 (WRN) S1141 Uniprot
Q14191 (WRN) S1292 Uniprot
Q14674-2 (ESPL1) S1615 Uniprot
Q14676 (MDC1) S513 Uniprot
Q14683 (SMC1A) S957 Uniprot
Q66K89 (E4F1) S356 Uniprot
Q6PGQ7 (BORA) T501 Uniprot
Q7Z6Z7 (HUWE1) S2391 Uniprot
Q8IUQ4 (SIAH1) S19 Uniprot
Q8IX01 (SUGP2) T7 Uniprot
Q8N163 (CCAR2) T454 Uniprot
Q8N4S0 (CCDC82) S154 Uniprot
Q8TAQ2 (SMARCC2) S969 Uniprot
Q8WVB6 (CHTF18) S74 Uniprot
Q8WXE1 (ATRIP) S68 Uniprot
Q8WXE1 (ATRIP) S72 Uniprot
Q92878 (RAD50) S635 Uniprot
Q92888-2 (ARHGEF1) S866 Uniprot
Q96RL1 (UIMC1) S205 Uniprot
Q96SD1 (DCLRE1C) S516 Uniprot
Q96SD1 (DCLRE1C) S645 Uniprot
Q99708 (RBBP8) T859 Uniprot
Q9BUG6 (ZSCAN5A) T422 Uniprot
Q9BXW9 (FANCD2) T596 Uniprot
Q9BXW9 (FANCD2) T608 Uniprot
Q9BXW9 (FANCD2) T691 Uniprot
Q9BXW9 (FANCD2) S717 Uniprot
Q9H6E5 (TUT1) S750 Uniprot
Q9NS23-4 (RASSF1) S61 Uniprot
Q9NS23-2 (RASSF1) S131 Uniprot
Q9NZC9 (SMARCAL1) S652 Uniprot
Q9NZN8 (CNOT2) S101 Uniprot
Q9UBU7 (DBF4) T449 Uniprot
Q9UBU7 (DBF4) S502 Uniprot
Q9UBU7 (DBF4) S539 Uniprot
Q9UER7 (DAXX) S424 Uniprot
Q9UFW8 (CGGBP1) S164 Uniprot
Q9UH99 (SUN2) S12 Uniprot
Q9Y248 (GINS2) S182 Uniprot
Q9Y253 (POLH) S601 Uniprot
Q9Y285 (FARSA) S301 Uniprot

Research Backgrounds

Function:

Serine/threonine protein kinase which activates checkpoint signaling upon genotoxic stresses such as ionizing radiation (IR), ultraviolet light (UV), or DNA replication stalling, thereby acting as a DNA damage sensor. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates BRCA1, CHEK1, MCM2, RAD17, RPA2, SMC1 and p53/TP53, which collectively inhibit DNA replication and mitosis and promote DNA repair, recombination and apoptosis. Phosphorylates 'Ser-139' of histone variant H2AX at sites of DNA damage, thereby regulating DNA damage response mechanism. Required for FANCD2 ubiquitination. Critical for maintenance of fragile site stability and efficient regulation of centrosome duplication.

PTMs:

Phosphorylated; autophosphorylates in vitro.

Subcellular Location:

Nucleus. Nucleus>PML body. Chromosome.
Note: Depending on the cell type, it can also be found in PML nuclear bodies. Recruited to chromatin during S-phase. Redistributes to discrete nuclear foci upon DNA damage, hypoxia or replication fork stalling.

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

Ubiquitous, with highest expression in testis. Isoform 2 is found in pancreas, placenta and liver but not in heart, testis and ovary.

Subunit Structure:

Interacts with ATRIP; forming a heterodimer with ATRIP. Binds to DNA, and to UV-damaged DNA with higher affinity. Interacts with RAD17, MSH2 and HDAC2. Present in a complex containing ATRIP and RPA-coated single-stranded DNA. Present in a complex containing CHD4 and HDAC2. Interacts with BCR-ABL after genotoxic stress. Interacts with EEF1E1. This interaction is enhanced by UV irradiation. Interacts with CLSPN and CEP164. Interacts with TELO2 and TTI1.

Family&Domains:

Belongs to the PI3/PI4-kinase family. ATM 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)

· Genetic Information Processing > Replication and repair > Fanconi anemia pathway.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

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

References

1). LncRNA OTUD6B-AS1 promotes paclitaxel resistance in triple negative breast cancer by regulation of miR-26a-5p/MTDH pathway-mediated autophagy and genomic instability. Aging, 2021 (PubMed: 34740994) [IF=5.2]
2). EBV encoded miRNA BART8-3p promotes radioresistance in nasopharyngeal carcinoma by regulating ATM/ATR signaling pathway. BIOSCIENCE REPORTS, 2019 (PubMed: 31471531) [IF=4.0]

Application: WB    Species: mouse    Sample: EBV-negative NPC cell

Figure 5. |EBV-miR-BART8-3p activates ATM/ATR. (A) Western blot analysis of p-ATM/ATM, p-ATR/ATR, and p-DNA-PKcs/DNA-PKcs in response to 0–12 h of irradiation at 2 Gy in the two groups.
3). OC-STAMP Overexpression Drives Lung Alveolar Epithelial Cell Type II Senescence in Silicosis. Oxidative Medicine and Cellular Longevity, 2021 (PubMed: 34426759)

Application: WB    Species: Mice    Sample: MLE-12 cells

Figure 3 Overexpression of OC-STAMP promotes ER stress and cellular senescence in MLE-12 cells. (a) Levels of p-PERK, p-EIF 2α, p-NF-κB, and p-IRE 1α in MLE-12 cells, measured by Western blotting. Data are presented as the mean ± SD; n = 3 per group; (b) SA-β-gal staining; (c) coexpression of PCNA and p21 in MLE-12 cells. Bar = 50 μm; (d) levels of p-ATM, p-ATR, p-p53, p21, and p16 in MLE-12 cells measured by Western blotting. Data are presented as the mean ± SD; n = 3 per group.

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