生长停滞DNA损伤可诱导蛋白α(GADD45a)活性蛋白

[ PROPERTIES ]

Source: Prokaryotic expression. Host: E. coli

Residues: Met1~Arg165

Tags: N-terminal His-tag

Purity: >95%

Endotoxin Level: <1.0EU per 1μg (determined by the LAL method). Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl

and 5% trehalose. Applications: Cell culture; Activity Assays. (May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 4.5

Predicted Molecular Mass: 22.2kDa

Accurate Molecular Mass: 22kDa as determined by SDS-PAGE reducing conditions.

[ USAGE ]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex. 

[ STORAGE AND STABILITY ]

Storage: Avoid repeated freeze/thaw cycles. 

Store at 2-8^oC for one month. Aliquot and store at -80^oC for 12 months.

Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37^oC for 48h, and no obvious degradation and precipitation wereobserved. The loss rate is less than 5% within the expiration date underappropriate storage condition. 

[ SEQUENCE ]

[ ACTIVITY ] 

Growth arrest and DNA-damage-inducible protein GADD45 alpha (GADD45a) is amember of the GADD45 family. It may affect PCNA interaction with some CDK(cell division protein kinase) complexes; stimulates DNA excision repair in vitroand inhibits entry of cells into S phase. In T-cells, functions as a regulator of p38MAPKs by inhibiting p88 phosphorylation and activity. Besides, Proliferating CellNuclear Antigen (PCNA) has been identified as an interactor of GADD45a, thus abinding ELISA assay was conducted to detect the interaction of recombinant ratGADD45a and recombinant rat PCNA. Briefly, GADD45a were diluted serially inPBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferredto PCNA-coated microtiter wells and incubated for 2h at 37℃. Wells were washedwith PBST and incubated for 1h with anti-GADD45a pAb, then aspirated andwashed 3 times. After incubation with HRP labelled secondary antibody, wellswere aspirated and washed 3 times. With the addition of substrate solution, wellswere incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wellsand read at 450nm immediately. The binding activity of GADD45a and PCNA wasshown in Figure 1, and this effect was in a dose dependent manner.

[ IDENTIFICATION ]

[ IMPORTANT NOTE ] 

The kit is designed for research use only, we will not be responsible for any issue if the kit was used in clinical diagnostic or any other procedures.

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