Application Note
IL1RA (interleukin-1 receptor antagonist) is an agent that binds to the cell surface interleukin-1 receptor (IL-1R), which would prevent IL-1 from intracellular signal transduction. Besides, mouse IL1R1 and rat IL1R1 share similarities in amino acid sequence with the identity of 85.6% . Thus, a binding ELISA assay was constructed to detect the association of recombinant mouse IL1RA with recombinant rat IL1R1. Briefly, IL1RA were diluted serially in PBS with 0.1% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to IL1R1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-IL1RA 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 IL1RA with IL1R1 was in a dose dependent manner.
Observed MW
20 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; Arg27~Gln178 (NP_001034790.1)
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.