Application Note
Growth arrest and DNA-damage-inducible protein GADD45 alpha (GADD45a) is a member of the GADD45 family. It may affect PCNA interaction with some CDK (cell division protein kinase) complexes; stimulates DNA excision repair in vitro and inhibits entry of cells into S phase. In T-cells, functions as a regulator of p38 MAPKs by inhibiting p88 phosphorylation and activity. Besides, Proliferating Cell Nuclear Antigen (PCNA) has been identified as an interactor of GADD45a, thus a binding ELISA assay was conducted to detect the interaction of recombinant rat GADD45a and recombinant rat PCNA. Briefly, GADD45a were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to PCNA-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-GADD45a pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of GADD45a and PCNA was in a dose dependent manner.
Observed MW
22 kDa.
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Met1~Arg165
Expression System
E. coli
Purity
> 95%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Synonyms
growth arrest and DNA-damage-inducible, alpha , Ddit1 , Gadd45
Background
may mediate a delay in G2 to M cell cycle progression; may induce DNA repair [RGD, Feb 2006]
Database
Research Area