The d‐THP‐1 were obtained by culturing in RPMI medium in the presence of PMA for 48 hours in six‐well plates as described above. LPS (10 ng/mL) was added and media (100 μL) were collected every four hours during the 24 hours treatment. For u‐THP‐1 and d‐THP‐1 comparison study, cells were cultured in six‐well plate as described above and media (100 μL) were collected from both u‐THP‐1 and d‐THP‐1 treated with/without 10 ng/mL of LPS for 24 hours. CXCL12 protein levels in the media were determined using a commercially available human CXCL12/SDF‐1α ELISA kit according to the manufacturer's instructions (R&D Systems, Minneapolis, MN).
Human cxcl12 sdf 1α elisa kit
Manufactured by R&D Systems
Sourced in United States
The Human CXCL12/SDF-1α ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of CXCL12/SDF-1α levels in human cell culture supernatants, serum, and plasma samples.
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3 protocols using human cxcl12 sdf 1α elisa kit
1
Monocyte Differentiation and CXCL12 Response
2
CXCL12/SDF-1α Quantification by ELISA
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CXCL12/SDF-1α levels in the supernatant of mono- and co-cultured cells were measured by the human CXCL12/SDF-1α ELISA kit (R and D Systems, Minneapolis, MN, USA) using 100 μL conditioned cell medium per sample. The assay was carried out according to the manufacturer’s instructions. Each measurement was performed in duplicate and the mean values were used for statistical analysis. ELISA plates were read at a wavelength of 570 nm using Labsystems Multiscan MS 352 plate reader (Labsystems, Finland).
3
Quantifying EPCs and SDF-1α in AVM
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A total of 15 AVM patients were randomly selected for venous blood collection, along with 15 healthy controls. Flow cytometry technique was applied to analyze the percentage of EPCs in the total mononuclear cells as previously reported.11 (link) EPCs were defined as triple-positive signals for CD133, CD34, and KDR. Enzyme-linked immunosorbent assay (ELISA) was also employed to detect the level of SDF-1α in peripheral blood by using previously established protocols12 (link) and the human CXCL12/SDF-1α ELISA kit (R&D Systems, Minneapolis, MN, USA). A correlation analysis was then performed to identify the correlation between EPCs percentage and SDF-1α level in each individual sample.
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