Maternal blood was collected at 2, 3 and 24 h post-injection from the tail vein and plasma harvested [20 (link)]. Maternal plasma IL-6, TNFα and IL-1β concentration was determined using rat-specific ELISAs according to the manufacturer’s instructions (Abcam, Cambridge, U.K.; ab100772, ab100784 and ab100767, respectively).
Ab100767
Manufactured by Abcam
Sourced in United Kingdom
Ab100767 is a product offered by Abcam. It is a piece of lab equipment. No further details about its core function or intended use can be provided in a factual and unbiased manner.
Automatically generated - may contain errors
Lab products found in correlation
Ab100772, by Abcam (2 mentions)
Ab46070, by Abcam (2 mentions)
Ab119548, by Abcam (2 mentions)
Infinite 200 pro microplate reader, by Tecan (1 mentions)
Eon spectrophotometer, by Agilent Technologies (1 mentions)
Ab234570, by Abcam (1 mentions)
Ab213909, by Abcam (1 mentions)
Bms630, by Thermo Fisher Scientific (1 mentions)
Ab100785, by Abcam (1 mentions)
Transwell plate inserts, by Merck Group (1 mentions)
Ab119602, by Abcam (1 mentions)
6 protocols using ab100767
1
Maternal cytokine response after injection
2
Macrophage Cytokine Profiling from Biomaterial Conditioned Media
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F-BMC, BMC or fibrin were cultured in the control medium (DMEM, 10% FBS, 1% P/S). After 48 h, the media were harvested and centrifuged. The macrophages were cultured for 48 h, with the conditioned media of F-BMC, BMC or substrate. Then, macrophages were washed with PBS and cultured with standard medium (DMEM, 10% FBS, 1% P/S). After 48 h, the cytokines were quantified in the medium harvested from the educated macrophages. ELISA kits (IL-1β (ab100767; Abcam, Cambridge, UK), IL-6 (ab119548; Abcam, Cambridge, UK), TNF-alfa (ab46070; Abcam, Cambridge, UK)) were used according to the manufacturer’s instructions and were estimated as pg/mL. Absorbance was read on a Tecan Infinite 200 PRO Microplate Reader (Tecan Group, Mennedorf, Switzerland).
3
Measurement of Inflammatory Cytokines in Rat Samples
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Following deep anesthesia in rats, blood was obtained from the ocular venous plexus and allowed to stand at 4°C for 1 h. This was followed by centrifugation at 700 rpm, and the supernatant was collected for detection. The IL-6 and IL-1β levels in the clinical plasma samples, rat peripheral blood, and culture supernatant of PASMCs were measured by the reference to the instructions of rat IL-6 and IL-1β ELISA kits (ab234570 and ab100767, respectively; Abcam, Cambridge, UK). The value in the microplate at 450 nm was recorded and analyzed using an Eon spectrophotometer (BioTek Instruments, Winooski, VT, USA). The experiment was conducted in triplicate independently.
4
Quantifying IL-1β and IL-18 Levels
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5
Modulating Inflammatory Responses via Co-culture
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The co-culture experimental design is shown in Figure 7 A. HAPI rat microglial cells (SCC103, Sigma-Aldrich, Germany) were seeded in 6-well plates (1×105 cells each). To induce HAPI cell reactivity, we used 1 mg/mL of LPS diluted in complete medium for 4 h. OECs or siRNA OECs were seeded (1×105 cells each) in Transwell plate inserts (Millipore). HAPI cells were co-cultured with the OECs or siRNA OECs for another 24 h. To explore their roles in modulating inflammation, the culture supernatants were used for ELISA of pro-inflammatory cytokines, including TNFα (ab100785, Abcam, Cambridge, UK), IL-1β (ab100767, Abcam, Cambridge, UK), and IL-6 (ab100772, Abcam, Cambridge, UK).
6
Biomarkers of Kidney Injury Assessment
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Blood samples were collected via the abdominal aorta, and urine samples were collected via the metabolic cage. Creatinine (E02C0629; Blue Gene) and blood urea nitrogen (E02C0697; Blue Gene) in the serum were measured using enzyme-linked immunosorbent assay (ELISA) kits. Neutrophil gelatinase-associated lipocalin (NGAL) was measured using rat NGAL ELISA kits (ab119602; Abcam). S-Thrombomodulin was measured using the Rat Soluble thrombomodulin (sTM) ELISA kit (E02S0222; Blue Gene). Serum levels of tumor necrosis factor (TNF)-α (ab46070; Abcam), inter-leukin (IL)-1β (ab100767; Abcam), and IL-6 (ab119548; Abcam) were measured using a rat bioactive ELISA.
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