Application Note
IL2RA (Interleukin-2 receptor alpha chain ), also known as Tac antigen and as CD25, was initially identified as a 55kDa membrane glycoprotein which is capable of binding IL 2. Besides, mouse IL2 shares 75.1% AA sequence identity with rat IL2, suggesting the exist of cross-species activity. Thus we have conducted a binding ELISA assay to detect the interaction of recombinant rat IL2RA with recombinant mouse IL2. Briefly, IL2RA were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to IL2-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-IL2RA 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 IL2RA with IL2 was in a dose dependent manner.
Observed MW
37 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; Glu22~Lys236 (NP_032393.3)
Expression System
E. coli
Purity
> 90%
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.