- First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions. - If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer. - Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent. - Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised. - Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.
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DNA dependent protein kinase catalytic subunit
DNA PK catalytic subunit
DNA-dependent protein kinase catalytic subunit
DNA-PK catalytic subunit
DNAPK catalytic subunit
Hyper radiosensitivity of murine scid mutation, complementing 1
Protein Kinase DNA Activated Catalytic Polypeptide
功能Serine/threonine-protein kinase that acts as a molecular sensor for DNA damage. Involved in DNA nonhomologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination. Must be bound to DNA to express its catalytic properties. Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step. Required to protect and align broken ends of DNA. May also act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage. Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion. Also involved in modulation of transcription. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX, thereby regulating DNA damage response mechanism. Phosphorylates DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, SRF, XRCC1, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2. Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA. Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D.
翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR. Autophosphorylated on Thr-2609, Thr-2638 and Thr-2647. Thr-2609 is a DNA damage-inducible phosphorylation site (inducible with ionizing radiation, IR). Autophosphorylation induces a conformational change that leads to remodeling of the DNA-PK complex, requisite for efficient end processing and DNA repair. S-nitrosylated by GAPDH.
Western blot - DNA PKcs peptide - phospho S2056 (ab20406)
All lanes : Anti-DNA PKcs (phospho S2056) antibody - ChIP Grade (ab18192) at 1 µg/ml
Lane 1 : HeLa Gamma Irradiated Whole Cell Lysate Pack (ab13823) at 20 µg Lane 2 : 20ug untreated HeLa cell extract Lane 3 : HeLa Gamma Irradiated Whole Cell Lysate Pack (ab13823) at 20 µg with Human DNA PKcs (phospho S2056) peptide (ab20406) at 1 µg/ml Lane 4 : 20ug untreated HeLa cell extract with Human DNA PKcs (phospho S2056) peptide (ab20406) at 1 µg/ml Lane 5 : HeLa Gamma Irradiated Whole Cell Lysate Pack (ab13823) at 20 µg with Human DNA PKcs peptide (ab20407) at 1 µg/ml Lane 6 : 20ug untreated HeLa cell extract with Human DNA PKcs peptide (ab20407) at 1 µg/ml
Secondary Alexa Fluor Goat polyclonal to Rabbit IgG (700)
at 1/10000 dilution