Product Name:
DAPK-pS269
Product Number:
ab-pk590
Target Full Name: Mitogen-activated protein kinase kinase kinase 12
Target Alias: Dual leucine zipper bearing kinase; Dual leucine zipper kinase DLK; EC 2.7.11.25; Kinase DLK; Leucine-zipper protein kinase; M3K12; MAP3K12; MAPK-upstream kinase; MEKK12; MUK; ZPK; ZPKP1
Product Type Specific: DAPK protein kinase phosphosite-specific antibody
Antibody Code: PK590
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S269
Protein UniProt: Q12852
Protein SigNET: DAPK
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human DLK (ZPK) sequence peptide Cat. No.: PE-04ALS85
Antibody Immunogen Sequence: TKM(pS)FAG(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues T266 to G272; in the protein kinase catalytic domain activation T loop region between subdomains VII and VIII. S269 phosphorylation is predicted to be stimulatory for phosphotransferase activity. This is one of the minor in vivo phosphorylation sites in DLK (DAPK1) (≥2 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Very strong immunoreactivity of a target-sized protein by Western blotting in A431 cells. Very strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus Macaque | Dog | Rat | Mouse | Chicken | Frog | Zebra fish | Honey bee | Sea urchin.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 93-96 kDa.
Antibody Specificity: High
Antibody Cross Reactivity: No significant cross-reactive proteins detected in HepG2 cells and sea star oocytes, except for a ~76 kDa cross-reactive protein and phospho-CDK1 in germinal vesicle-positive, maturing sea star oocytes.
Related Product 1: DLK (266-272) pS269 antibody blocking peptide (Cat. No.: PE-04ALS85)
Related Product 2: MYL9 (12-23) KinSub - Myosin light chain 9 (A12-S23, human) peptide; PHKG protein kinase substrate peptide
Related Product 3: DAPKSubtide kinase substrate peptide (Cat. No.: PE-01BGZ95)
Related Product 4: ZIPtide KinSub kinase substrate peptide (Cat. No.: PE-01ANB95)
Related Product 5: DAPK1 S308 phosphorylation antibody (Cat. No.: AB-PK900)
Scientific Background: DLK (MAP3K12; DAPK) is a protein-serine/threonine kinase of the TKL group and MLK family. It is a protein kinase that has been noted as an upstream activator of the JNK (c-Jun N-terminal kinase) and p38 MAPK pathways. It features a catalytic kinase domain and a distinctive "dual leucine zipper" domain (two zipper motifs separated by a short spacer) near the C-terminus, which mediates dimerization and activation. Upon injury, DLK dimerizes via its leucine zipper domains and autophosphorylates. It binds to JNK-interacting proteins (JIPs), such as JIP1, JIP2, and JIP3, which help assemble the signaling module. DLK activates MKK4/7, which in turn activates JNK, leading to the phosphorylation of the transcription factor c-Jun. In vivo DLK has also been noted to phosphorylate beta-casein, myelin basic protein, and histone 1. In the developing nervous system, DLK is essential for proper axon tract formation and neuronal migration, often regulating microtubule stability via the JNK-dependent phosphorylation of targets like MAP1B and doublecortin. Upon axonal injury, DLK is rapidly activated in the damaged axon and serves as a sensor, triggering a retrograde signal that promotes regeneration (via STAT3 and JIP3) in the peripheral nervous system (PNS). In the central nervous system (CNS), when injury is severe, DLK mediates a "self-destruction" program (Wallerian degeneration) where it drives the degeneration of damaged axons and promotes neuronal cell death (apoptosis). Its activity is heavily regulated by dimerization, palmitoylation (which restricts it to transport vesicles), and ubiquitination-mediated degradation by the PHR family of E3 ligases (e.g., Phr1/Highwire/RPM-1). DLK is most abundantly expressed in neural tissues, specifically in differentiated neurons rather than proliferating cells. In the developing brain, it is found in the subventricular and intermediate zones. It is localized predominantly in axons and growth cones, often associated with microtubules and the Golgi apparatus. While necessary for regeneration, DLK also mediates neuronal loss in chronic neurodegenerative diseases such as Alzheimer's, Parkinson’s, and Amyotrophic Lateral Sclerosis (ALS). Since overactivation leads to neurodegeneration, several small-molecule inhibitors have been developed (e.g., GNE-3511, GNE-8505, GDC-0134, and DN-1289) to prevent neuronal damage and inhibit the stress response, and these show potential in alleviating symptoms of Alzheimer's, glaucoma, and ALS. This description may include information annotated by UniProt and/or Google AI.