Human g csf instant elisa kit

Pharmacokinetic profiles
of G-CSF and G-CSF conjugates (cPEG_20k-Nter/K41-G-CSF,
PEG_20k-Nter-G-CSF, and PEG_20k-K41-G-CSF) were
determined in female Sprague–Dawley rats weighing between 150
g and 160 g (3 animals per group). Samples were prepared in PBS pH
7.4 and were intravenously injected at the dose of 100 μg/kg
(G-CSF equivalent) in the lateral tail vein. Anesthesia was performed
with 5% isoflurane gas mixed with O₂ in enclosed cages.
Blood samples were collected on anesthetized rats by incision from
the tail at predetermined time points and centrifuged at 1500g for 20 min. G-CSF equivalent concentrations in serum were
quantified using a Human G-CSF Instant ELISA Kit (Life Technologies)
by using the corresponding conjugates as standard. Pharmacokinetic
data were elaborated using 2.0 PkSolver software by applying a bi-compartmental
model. For each sample, a dedicated calibration curve was built using
the testing conjugate.

The protein rhG-CSF was a kind gift of Sandoz
(Ljubljana, Slovenia). PEG-aldehyde 20 kDa and PEG-NH₂ 20
kDa were purchased from NOF Corporation (Tokyo, Japan). The H₂N-PEG-COOH 20 kDa was purchased from JenKem Technology USA,
Inc. (Plano, TX). Microbial TGase (mTGase), of Streptomyces
mobaraensis origin (ACTIVA M), was provided by Ajinomoto
Co. (Tokyo, Japan). Carbobenzoxy-l-glutaminyl-glycine and
all of the chemicals and solvents were purchased from Merk (Darmstadt,
Germany). Trypsin and Glu-C of sequencing grade were obtained by Thermo
Fisher Scientific (Waltham, MA). Human G-CSF Instant ELISA Kit was
purchased from Life Technologies (Waltham, MA). Mini-PROTEAN TGX precast
gels 4–15% for SDS-PAGE were obtained from Bio-Rad (Milan,
Italy).