1.25 or 2.5 μg of Chi3l1 siRNA (Bioneer) or EGFP siRNA (Bioneer) was incubated with 35 or 70 μg dNP2-HA2 peptide for 30 min at RT, in the dark. Free siRNA or Peptide-siRNA complexes were intranasally administered to mice, which were sacrificed every 24 h for 3 days. Lung tissue was disrupted in T-PER Tissue Protein Extraction Reagent (Thermo Fisher Scientific) for protein lysates or in TRIzol for RNA extraction. Chi3l1 protein in the lung was measured by Chi3l1 ELISA kit (R&D Systems). In the lung metastasis model, 5 × 105 B16F10 cells were intravenously injected to WT mice, and then 2.5 μg of siRNA + 70 μg of dNP2-HA2 peptide complex was intranasally administered to anesthetized mice with zoletil + Rompun for twice per day with 10 h interval for every other day until day 14. At 14 days after melanoma injection, mice were sacrificed and analyzed.
Chi3l1 elisa kit
Manufactured by R&D Systems
Sourced in United States
The Chi3l1 ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) for the measurement of Chi3l1 in human samples. The kit utilizes a specific antibody coated onto a microplate to capture Chi3l1 from samples, which is then detected with a secondary antibody conjugated to an enzyme. The resulting color change is proportional to the amount of Chi3l1 present in the sample.
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2 protocols using chi3l1 elisa kit
1
Intranasal Delivery of Chi3l1 siRNA-Peptide Complex
2
Quantifying CHI3L1 in MS and Controls
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ELISA analysis of CHI3L1 levels ELISA samples were derived from human serum and cerebrospinal fluid (CSF) of patients with MS and healthy controls, as well as lysates of freshly dissected mouse hippocampal tissues. Blood samples were collected into biochemical tubes and centrifuged at 8000 rpm for 15 min at 4 C. Plasma was collected and stored at À80 C until analysis. The determination of CHI3L1 in serum and CSF was conducted using a commercial CHI3L1 ELISA kit (R&D Systems, MN, USA) according to the manufacturer's instructions. Mouse hippocampal tissues were homogenized in RIPA buffer (P0013, Beyotime) and centrifuged at 12,000g for 30 min at 4 C. The concentration of CHI3L1 was measured with a mouse ELISA kit (MEIMIAN, MM-44752M1) at 450 nm. All assays were performed in duplicate.
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