A high-sensitivity IL-6 ELISA kit was performed in accordance to the manufacturer’s instructions (BMS625, Invitrogen; Thermo Fisher Scientific, Inc.). In brief, washing buffer, assay buffer, and standards were prepared and reconstituted with ddH2O and serial dilutions of the standard curve were externally prepared using assay buffer. A pre-coated 96 micro-well plate was used and read at 450nm. Results were calculated according to the best-fit standard curve.
Bms625
Manufactured by Thermo Fisher Scientific
Sourced in United States
The BMS625 is a laboratory instrument designed for automated sample preparation. It is capable of performing various liquid handling tasks, including pipetting, dilution, and sample transfer. The BMS625 is intended to improve efficiency and accuracy in the laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Bms630, by Thermo Fisher Scientific (6 mentions)
Bms629, by Thermo Fisher Scientific (6 mentions)
Bms622, by Thermo Fisher Scientific (4 mentions)
Krc3011, by Thermo Fisher Scientific (2 mentions)
Ab100768, by Abcam (2 mentions)
Ab100785, by Abcam (2 mentions)
Bms634, by Thermo Fisher Scientific (2 mentions)
Varioskan lux, by Thermo Fisher Scientific (1 mentions)
Cobas 8000 e602, by Roche (1 mentions)
Synergy 2, by Agilent Technologies (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Cat kit, by Nanjing Jiancheng (1 mentions)
Sod kit, by Nanjing Jiancheng (1 mentions)
Mda kit, by Nanjing Jiancheng (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Ab61224, by Abcam (1 mentions)
Elr leptin 1, by RayBiotech (1 mentions)
Milliplex analyst software, by Merck Group (1 mentions)
Milliplex magnetic bead panel kits, by Thermo Fisher Scientific (1 mentions)
21 protocols using bms625
1
Sensitive IL-6 ELISA Protocol
2
Quantifying Cytokine Levels in SAH
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At 24 h and 72 h post-SAH, rats were anesthetized and transcardially perfused with 250 mL PBS, the left hemisphere was harvested immediately and homogenized in 0.9% normal saline at 200 mg/mL and centrifuged at 12,000×g for 20 min. The supernatant was collected and stored in − 80 °C until further used. The concentrations of TNF-α, interleukin (IL)-6, IL-1β, and IL-10 in brain tissue lysates were analyzed using commercial ELISA kits (BMS625, BMS630, and BMS629, Invitrogen) according to the manufacturer’s instructions. The final concentration of cytokines was interpolated from the determined standard curve of absorbance.
3
Serum Cytokine Profile after SAH
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At 24 h after SAH, rats were deeply anesthetized and serum samples from the left hemisphere were obtained and stored at −80°C until use. The concentrations of IL-10, IL-1β, and IL-18 in brain tissue lysates were analyzed using commercial ELISA kits (BMS625, BMS630, and BMS629, Invitrogen) according to the manufacturer's instructions. The final concentration of cytokines was obtained from the determined standard curve of absorbance.
4
Quantification of Inflammatory Cytokines
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The samples of brain tissues and cultured cells were prepared as WB. The protein levels of TNF-α (#KRC3011, Invitrogen), IL-1β (ab100768, Abcam), and IL-6 (#BMS625, Invitrogen) were determined by ELISA according to the manufacturer's instructions. The standards were run in duplicate and samples in triplicate. Absorbance was measured at 450 nm using the Bio-Rad DC. Cytokine concentrations were normalized to the total protein content and reported as pg/mg of total protein ± S.E.M.
5
Cytokine Profiling in Rat SAH
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At 24 h post-SAH, rats were anesthetized and transcardially perfused with 250 mL pre-cooled PBS (0.1 M, pH 7.4). Brains were then removed and rapidly divided into the left and right hemisphere; the left hemisphere was frozen in an ultra-low temperature refrigerator, homogenized in PBS (0.1 M, pH 7.4) at 200 mg/mL, and centrifuged at 12,000 g for 15 min. The liquid supernatant was collected and stored at −80 °C until further use. The concentrations of TNF-α, IL-1β, IL-17and IL-18 were analyzed using specific ELISA kits (BMS625, BMS630, and BMS629, respectively, Invitrogen) according to the manufacturer’s instructions. The concentration of cytokines was expressed by the intensity of absorbance measured by spectrometry on a microplate reader (Varioskan Lux, Thermo Scientific). Results were expressed in picograms per milliliter (pg/ml).
6
Plasma Cytokine Analysis after MI
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Two weeks after MI and patch treatment, blood samples were collected and transferred to EDTA tubes. The blood samples were centrifuged for 10 min at 25 °C, and plasma was stored at −80 °C until assay. Tumor necrosis factor (TNF-α) and interleukin-6 (IL-6) in plasma were quantified using ELISA kits (BMS622 and BMS625, Invitrogen, Waltham, MA, USA), according to the manufacturer's instructions.
7
Quantifying Brain Inflammation Biomarkers
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Interleukin-6 (IL-6), interleukin 1β (IL-1β), and tumor necrosis factor-alpha (TNF-α) were colorimetrically detected in the brain tissue according to the manufacturers' instructions based on the sandwich and competitive ELISA technique using commercially available kits for rat IL-6 (BMS625), (IL-1β) (BMS630), and TNF-α (BMS622), (Invitrogen, Thermo Fisher Scientific, MI, USA). IL-6, IL-1β, and TNF-α concentrations were measured in brain homogenate as pg/ml at 450 nm by interpolation from a standard curve via a micro-plate ELISA reader (Sorin Biomedica SpA., Milan, Italy) with a detection limit from 31.3–2000 pg/ml, 31.3–2000 pg/ml, and 39.1–2500 pg/ml, respectively (Jiang et al. 2021 (link)).
8
Measuring Serum and Tissue Biomarkers
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The serum LDH level was determined using a cobas 8000 e602 modular automatic
analyzer (Roche Diagnostics, Mannheim, Germany). Serum and intestinal tissue
levels of TNFα were measured using enzyme-linked immunosorbent assay (ELISA)
kits (bms622, bms622TWO, and bms622TEN; Affymetrix eBioscience, San Diego, CA,
USA). Serum and tissue levels of IL-6 were determined using ELISA kits (bms625,
bms625TWO, and bms625TEN; Affymetrix eBioscience). Serum and tissue levels of
IL-1β were determined using ELISA kits (bms630 and bms630TEN; Affymetrix
eBioscience). Serum and tissue levels of MPO were determined using ELISA kits
(201-11-0575; Shanghai Sunred Biological Technology Co., Ltd., Shanghai, China).
Serum and tissue levels of MDA were determined using ELISA kits (201-11-0157;
Shanghai Sunred Biological Technology Co., Ltd.).
analyzer (Roche Diagnostics, Mannheim, Germany). Serum and intestinal tissue
levels of TNFα were measured using enzyme-linked immunosorbent assay (ELISA)
kits (bms622, bms622TWO, and bms622TEN; Affymetrix eBioscience, San Diego, CA,
USA). Serum and tissue levels of IL-6 were determined using ELISA kits (bms625,
bms625TWO, and bms625TEN; Affymetrix eBioscience). Serum and tissue levels of
IL-1β were determined using ELISA kits (bms630 and bms630TEN; Affymetrix
eBioscience). Serum and tissue levels of MPO were determined using ELISA kits
(201-11-0575; Shanghai Sunred Biological Technology Co., Ltd., Shanghai, China).
Serum and tissue levels of MDA were determined using ELISA kits (201-11-0157;
Shanghai Sunred Biological Technology Co., Ltd.).
9
Quantifying IL-6 Levels in Biospecimens
10
Inflammatory Cytokine Modulation in LPS-Induced Sepsis
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The animals were divided into four groups randomly (eight animals in each group). The control group received vehicle (10 ml/kg of physiological saline) intraperitoneally (i.p.), while 10, 20, or 40 mg/kg of fluoxetine (i.p.) was injected to the fluoxetine groups. LPS (10 mg/kg, i.p.) was injected 30 min after drug administration. Blood sampling was carried out via cardiac puncture under anesthesia (40 mg/kg of pentobarbital, i.p.) 90 min following the injection of LPS. After coagulation, blood was centrifuged at 2000 g for 20 min to produce serum specimens. The serum levels of IL-1β, IL-6, and TNF-α were measured using commercial ELISA kits (Cat. No., IL-1β: BMS630; IL-6: BMS625; and TNF-α: KRC3011), based on the manufacturer’s instructions (eBioscience, San Diego, CA, USA). A microplate reader (BioRad, USA) was used to read absorbance at 450 nm.
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