Product: DAPK1 Antibody
Catalog: DF8030
Description: Rabbit polyclonal antibody to DAPK1
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 160kDa; 160kD(Calculated).
Uniprot: P53355
RRID: AB_2841398

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 100ul $280 In stock
 200ul $350 In stock

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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(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
DAPK1 Antibody detects endogenous levels of total DAPK1.
RRID:
AB_2841398
Cite Format: Affinity Biosciences Cat# DF8030, RRID:AB_2841398.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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

DAK1; DAP K1; DAP kinase 1; DAPK 1; DAPK; DAPK1; DAPK1_HUMAN; Death Associated Protein Kinase 1; Death-associated protein kinase 1; DKFZp781I035;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P53355 DAPK1_HUMAN:

Isoform 2 is expressed in normal intestinal tissue as well as in colorectal carcinomas.

Sequence:
MTVFRQENVDDYYDTGEELGSGQFAVVKKCREKSTGLQYAAKFIKKRRTKSSRRGVSREDIEREVSILKEIQHPNVITLHEVYENKTDVILILELVAGGELFDFLAEKESLTEEEATEFLKQILNGVYYLHSLQIAHFDLKPENIMLLDRNVPKPRIKIIDFGLAHKIDFGNEFKNIFGTPEFVAPEIVNYEPLGLEADMWSIGVITYILLSGASPFLGDTKQETLANVSAVNYEFEDEYFSNTSALAKDFIRRLLVKDPKKRMTIQDSLQHPWIKPKDTQQALSRKASAVNMEKFKKFAARKKWKQSVRLISLCQRLSRSFLSRSNMSVARSDDTLDEEDSFVMKAIIHAINDDNVPGLQHLLGSLSNYDVNQPNKHGTPPLLIAAGCGNIQILQLLIKRGSRIDVQDKGGSNAVYWAARHGHVDTLKFLSENKCPLDVKDKSGEMALHVAARYGHADVAQLLCSFGSNPNIQDKEEETPLHCAAWHGYYSVAKALCEAGCNVNIKNREGETPLLTASARGYHDIVECLAEHGADLNACDKDGHIALHLAVRRCQMEVIKTLLSQGCFVDYQDRHGNTPLHVACKDGNMPIVVALCEANCNLDISNKYGRTPLHLAANNGILDVVRYLCLMGASVEALTTDGKTAEDLARSEQHEHVAGLLARLRKDTHRGLFIQQLRPTQNLQPRIKLKLFGHSGSGKTTLVESLKCGLLRSFFRRRRPRLSSTNSSRFPPSPLASKPTVSVSINNLYPGCENVSVRSRSMMFEPGLTKGMLEVFVAPTHHPHCSADDQSTKAIDIQNAYLNGVGDFSVWEFSGNPVYFCCYDYFAANDPTSIHVVVFSLEEPYEIQLNQVIFWLSFLKSLVPVEEPIAFGGKLKNPLQVVLVATHADIMNVPRPAGGEFGYDKDTSLLKEIRNRFGNDLHISNKLFVLDAGASGSKDMKVLRNHLQEIRSQIVSVCPPMTHLCEKIISTLPSWRKLNGPNQLMSLQQFVYDVQDQLNPLASEEDLRRIAQQLHSTGEINIMQSETVQDVLLLDPRWLCTNVLGKLLSVETPRALHHYRGRYTVEDIQRLVPDSDVEELLQILDAMDICARDLSSGTMVDVPALIKTDNLHRSWADEEDEVMVYGGVRIVPVEHLTPFPCGIFHKVQVNLCRWIHQQSTEGDADIRLWVNGCKLANRGAELLVLLVNHGQGIEVQVRGLETEKIKCCLLLDSVCSTIENVMATTLPGLLTVKHYLSPQQLREHHEPVMIYQPRDFFRAQTLKETSLTNTMGGYKESFSSIMCFGCHDVYSQASLGMDIHASDLNLLTRRKLSRLLDPPDPLGKDWCLLAMNLGLPDLVAKYNTSNGAPKDFLPSPLHALLREWTTYPESTVGTLMSKLRELGRRDAADFLLKASSVFKINLDGNGQEAYASSCNSGTSYNSISSVVSR

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

PTMs - P53355 As Substrate

Site PTM Type Enzyme
T15 Phosphorylation
Y39 Phosphorylation
S57 Phosphorylation
T112 Phosphorylation
S289 Phosphorylation Q15418 (RPS6KA1) , P51812 (RPS6KA3)
K295 Acetylation
K297 Acetylation
K306 Ubiquitination
S308 Phosphorylation P53355 (DAPK1)
S313 Phosphorylation P53355 (DAPK1)
S319 Phosphorylation
S321 Phosphorylation
S324 Phosphorylation
S326 Phosphorylation
S333 Phosphorylation
T336 Phosphorylation
K410 Ubiquitination
K435 Ubiquitination
Y490 Phosphorylation P12931 (SRC)
Y491 Phosphorylation P12931 (SRC)
S606 Phosphorylation
K691 Ubiquitination
S724 Phosphorylation
S725 Phosphorylation
S728 Phosphorylation
S734 Phosphorylation P27361 (MAPK3) , P28482 (MAPK1)
S762 Phosphorylation
K875 Ubiquitination
K906 Ubiquitination
K912 Ubiquitination
K939 Ubiquitination
T1053 Phosphorylation
Y1060 Phosphorylation
S1115 Phosphorylation
T1266 Phosphorylation
S1267 Phosphorylation
T1269 Phosphorylation

PTMs - P53355 As Enzyme

Substrate Site Source
O14950 (MYL12B) S20 Uniprot
O43293 (DAPK3) T299 Uniprot
O43293 (DAPK3) S309 Uniprot
O43293 (DAPK3) S311 Uniprot
O43293 (DAPK3) S312 Uniprot
O43293 (DAPK3) S318 Uniprot
O43293 (DAPK3) S326 Uniprot
P04637 (TP53) T18 Uniprot
P04637 (TP53) S20 Uniprot
P04637 (TP53) S269 Uniprot
P09493 (TPM1) S283 Uniprot
P19105 (MYL12A) S19 Uniprot
P25205 (MCM3) S160 Uniprot
P38936 (CDKN1A) S146 Uniprot
P40429 (RPL13A) S77 Uniprot
P53355 (DAPK1) S308 Uniprot
P53355 (DAPK1) S313 Uniprot
P62753 (RPS6) S235 Uniprot
Q00613 (HSF1) S230 Uniprot
Q13224 (GRIN2B) S1303 Uniprot
Q13526 (PIN1) S71 Uniprot
Q14457 (BECN1) T119 Uniprot
Q16623 (STX1A) S188 Uniprot
Q96RR4 (CAMKK2) S511 Uniprot

Research Backgrounds

Function:

Calcium/calmodulin-dependent serine/threonine kinase involved in multiple cellular signaling pathways that trigger cell survival, apoptosis, and autophagy. Regulates both type I apoptotic and type II autophagic cell deaths signal, depending on the cellular setting. The former is caspase-dependent, while the latter is caspase-independent and is characterized by the accumulation of autophagic vesicles. Phosphorylates PIN1 resulting in inhibition of its catalytic activity, nuclear localization, and cellular function. Phosphorylates TPM1, enhancing stress fiber formation in endothelial cells. Phosphorylates STX1A and significantly decreases its binding to STXBP1. Phosphorylates PRKD1 and regulates JNK signaling by binding and activating PRKD1 under oxidative stress. Phosphorylates BECN1, reducing its interaction with BCL2 and BCL2L1 and promoting the induction of autophagy. Phosphorylates TSC2, disrupting the TSC1-TSC2 complex and stimulating mTORC1 activity in a growth factor-dependent pathway. Phosphorylates RPS6, MYL9 and DAPK3. Acts as a signaling amplifier of NMDA receptors at extrasynaptic sites for mediating brain damage in stroke. Cerebral ischemia recruits DAPK1 into the NMDA receptor complex and it phosphorylates GRINB at Ser-1303 inducing injurious Ca(2+) influx through NMDA receptor channels, resulting in an irreversible neuronal death. Required together with DAPK3 for phosphorylation of RPL13A upon interferon-gamma activation which is causing RPL13A involvement in transcript-selective translation inhibition.

Isoform 2 cannot induce apoptosis but can induce membrane blebbing.

PTMs:

Ubiquitinated by the BCR(KLHL20) E3 ubiquitin ligase complex, leading to its degradation by the proteasome.

Removal of the C-terminal tail of isoform 2 (corresponding to amino acids 296-337 of isoform 2) by proteolytic cleavage stimulates maximally its membrane-blebbing function.

In response to mitogenic stimulation (PMA or EGF), phosphorylated at Ser-289; phosphorylation suppresses DAPK1 pro-apoptotic function. Autophosphorylation at Ser-308 inhibits its catalytic activity. Phosphorylation at Ser-734 by MAPK1 increases its catalytic activity and promotes cytoplasmic retention of MAPK1. Endoplasmic-stress can cause dephosphorylation at Ser-308.

Subcellular Location:

Cytoplasm. Cytoplasm>Cytoskeleton.
Note: Colocalizes with MAP1B in the microtubules and cortical actin fibers.

Cytoplasm. Cytoplasm>Cytoskeleton.

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

Isoform 2 is expressed in normal intestinal tissue as well as in colorectal carcinomas.

Subunit Structure:

Interacts with KLHL20. Interacts (via death domain) with MAPK1 and MAPK3. Interacts with MAP1B (via N-terminus). Interacts with PRKD1 in an oxidative stress-regulated manner. Interacts with PIN1, PDCD6, BECN1, TSC2 and STX1A. Interacts (via kinase domain) with DAPK3 (via kinase domain). Interacts with GRINB (By similarity). Interacts (via death domain) with UNC5B (via death domain). Interacts with UNC5C (via death domain).

Family&Domains:

The autoinhibitory domain sterically blocks the substrate peptide-binding site by making both hydrophobic and electrostatic contacts with the kinase core.

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

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Bladder cancer.   (View pathway)

References

1). PTBP1 knockdown promotes neural differentiation of glioblastoma cells through UNC5B receptor. Theranostics, 2022 (PubMed: 35664063) [IF=12.4]

Application: WB    Species: Human    Sample: U251 cells

Figure 7 A DAPK1 inhibitor prevents the PTBP1 knockdown induced reprogramming. (A-E) Western blot study of NTN1, DAPK1, P-DAPK1(Ser308), P-P53(ser20), and caspase 3(p17) protein in U251 cells infected with sh-Luci and sh-PTBP1 for 3 and 7 days. (n = 3). As an internal reference protein, GAPDH was used. (F) Activity of PP2A during reprogramming. (n = 3). (G-I) TUJ1 and KI67 positive rates of six distinct groups constituted of lentivirus (sh-Luci or sh-PTBP1) and TC-DAPK6 (vehicle, 100 nM, 250 nM, and 500 nM; nine random fields from triplicate samples were recorded for quantification; TUJ1+ (%) = TUJ1+ M-cherry+/M-cherry+; KI67+ (%) = KI67+ M-cherry+/M-cherry+; M-cherry+ cells = 156-553 for each condition). The data are presented as mean ± SD. *P < 0.05, *** P < 0.001 vs. sh-Luci-3d or sh-PTBP1 + vehicle group. Dpi (d): days post infection; ND: not detected; NS: no significance. Scale: 100 µm.

2). DAPK1 Interacts with the p38 Isoform MAPK14, Preventing Its Nuclear Translocation and Stimulation of Bone Marrow Adipogenesis. STEM CELLS, 2022 (PubMed: 35403694) [IF=5.2]

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