After E2 treatment and N. meningitidis isolate infection, both cell types were further cultured for 48 h. Cytokines were detected using ELISA. The interleukin-6 (IL-6) level was detected using Human IL-6 ELISA Set (#555220, BD Bioscience, San Diego, CA, United States). The release of interleukin-8 (IL-8) was detected using a Human IL-8 ELISA Kit II (#550999, BD Bioscience). The TNF-α level was detected using a Human TNF ELISA Kit II (#550610, BD Bioscience). The cytokines mentioned above were all detected according to the manufacturer’s instructions and run in triplicate. The above-mentioned cytokine levels released by non-infected HBMECs and MECs were measured as controls.
Human il 8 elisa kit 2
Manufactured by BD
Sourced in United States
The Human IL-8 ELISA Kit II is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of human interleukin-8 (IL-8) levels in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab23738, by Abcam (2 mentions)
Ab9385, by Abcam (2 mentions)
M0873, by Agilent Technologies (2 mentions)
Human il 6 elisa set, by BD (1 mentions)
Human tnf elisa kit 2, by BD (1 mentions)
Resiquimod, by InvivoGen (1 mentions)
Htlr7, by InvivoGen (1 mentions)
Htlr8, by InvivoGen (1 mentions)
Lipofectin, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by PAN Biotech (1 mentions)
Fetal bovine serum (fbs), by PAN Biotech (1 mentions)
6 protocols using human il 8 elisa kit 2
1
E2 and N. meningitidis Cytokine Profiling
2
Measuring IL-8 Secretion in Co-Cultures
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After 24-h incubation, co-culture media were sampled and centrifuged at 16,000 g, and supernatants collected. AGS cell IL-8 secretion was measured by an enzyme-linked immunosorbent assay using the Human IL-8 ELISA Kit II (BD Biosciences, San Jose CA), in accordance with the manufacturer’s instructions.
3
Quantification of IL-8 and HGF in AML
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Supernatant of primary AML cells (106/ml) or AML cell lines KG-1a and OCI-AML-3 (2.0 × 105/ml) were collected after 48 h of in vitro cultivation at the indicated oxygen concentration by centrifugation (10 min, 2000 rpm). For long term cultures cell lines were pre-cultured under hypoxic conditions for 8 days prior to this treatment. IL-8 and HGF protein in the supernatant was quantified with the human IL-8 ELISA Kit II (BD Biosciences, San Jose, USA) and human HGF ELISA (R&D Systems, Minneapolis, USA) according to the manufacturer’s instructions. For quantification of IL-8 protein in patients with AML, serum samples were collected during routine surveillance at the indicated time points and spun down at 2000 rpm for 10 minutes. Serum was stored at −80 °C and IL-8 quantification was performed by ELISA as described.
4
Comprehensive Protein Analysis Techniques
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Western blotting analyses were performed using standard protocols. Antibodies used were as follow: anti-FOXA1 (ab23738, Abcam), anti-NSE (M0873, DAKO), anti-pERK1/2 (4370S, cell signaling), anti-ERK1/2 (0192S, cell signaling), anti-AR (06-680, Millipore), and anti-GAPDH (ab9385, Abcam). ChIP was performed as previously described 54 (link). Antibodies used include anti-FOXA1 (ab23738, Abcam), anti-RNA PolII p-Ser5 (04-1572, Millipore), and anti-H3K4me3 (04-745, Millipore). ELISA was performed using Human IL-8 ELISA kit II following manufacture’s protocol (550999, BD Bioscience).
5
Assaying TLR7/8 Activation and IL-8 Response
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Human TLR-transformed HEK293 cells are an established system to analyse tRNA-induced stimulation of human TLR7 and TLR8 and the IL-8 response was monitored as described in28 (link). In brief, HEK293XL cells stably transfected with hTLR7 and hTLR8 (Invivogen) were seeded in 96-well cell culture plates at 5 × 104 cells per well and cultured in DMEM (Pan Biotech) supplemented with 10% FBS (Pan Biotech) and 2 mM l -glutamine (Thermo Fisher Scientific). Twenty-four hours later, cells were transfected with in vitro-transcribed tSA2T5 (1 µg ml−1) or resiquimod (R848; Invivogen, 1 µg ml−1) as control activators of the TLR7–TLR8 signalling pathway55 (link) using Lipofectin (Thermo Fisher Scientific). After 3 h, medium was replaced and 16 h post-transfections, interleukin IL-8 in supernatants was measured using human IL-8 ELISA Kit II (BD Biosciences) according to the manufacturer’s instructions.
6
Comprehensive Protein Analysis Techniques
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