Serum was harvested from blood. Tissues (liver and spleen) were homogenized in RIPA buffer (1:10 time dilution; w/v). The lysate was centrifuged at 16,000 × g for 6 min at 4 °C and the supernatant was collected. Serum and tissue supernatant were diluted by diluent buffer and subjected to mouse ferritin ELISA (Abcam ab157713) or rat ferritin ELISA (Abcam ab157732) according to the manufacturer’s instructions.
Ab157713
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab157713 is a lab equipment product manufactured by Abcam. It serves a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
I 536, by Leinco Technologies (2 mentions)
T 703, by Leinco Technologies (2 mentions)
I 1190, by Leinco Technologies (2 mentions)
Ab208348, by Abcam (1 mentions)
Ab222503, by Abcam (1 mentions)
Dri chem slide, by Fujifilm (1 mentions)
Enzychrom free fatty acid assay kit, by BioAssay Systems (1 mentions)
Hdl c, by Fujifilm (1 mentions)
7 protocols using ab157713
1
Ferritin ELISA Quantification
2
Serum Biomarker Analysis Protocol
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3
Plasma IL-18 and Ferritin Quantification
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Sandwich ELISA was used to establish plasma concentrations of IL-18 (Invitrogen, Thermofisher, BMS618–3) and ferritin (Abcam, ab157713). ELISA protocol was carried out according to manufacturer’s instructions. ELISA was read at 450–570nm according to manufacturer’s instructions.
4
Cytokine-Induced Mortality in Mice
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Age- and gender-matched 6- to 8-week-old mice of the indicated genotypes were injected intraperitoneally with 10 μg TNF-α alone, 20 μg IFN-γ alone, or TNF-α + IFN-γ diluted in DPBS. Animals were under permanent observation, and survival was assessed every 30 min. Blood was collected 5 h after cytokine injection. Blood composition was analyzed using an automated hematology analyzer. Serum LDH, ALT, AST, blood urea nitrogen (BUN), and ferritin were analyzed by colorimetry using respective kits (LDH, REF#A11A01824; ALT, REF#A11A01627; AST, REF#A11A01629; and BUN, REF#A11A01641, all from HORIBA; and ferritin, ab157713, Abcam) according to the manufacturer’s instructions.
For neutralizing antibody treatment, age- and gender-matched 6- to 8-week-old WT mice were administered intraperitoneally 200 μL of DPBS containing 500 μg of isotype control (Leinco Technologies, Inc., I-536) (n = 10) or 500 μg of neutralizing antibody against TNF-α (Leinco Technologies, Inc., T-703) plus 500 μg of neutralizing antibody against IFN-γ (Leinco Technologies, Inc., I-1190) (n = 14) 12 h before i.p. injection of TNF-α and IFN-γ. Mice were monitored for survival. The results were pooled from 2 independent experiments.
For neutralizing antibody treatment, age- and gender-matched 6- to 8-week-old WT mice were administered intraperitoneally 200 μL of DPBS containing 500 μg of isotype control (Leinco Technologies, Inc., I-536) (n = 10) or 500 μg of neutralizing antibody against TNF-α (Leinco Technologies, Inc., T-703) plus 500 μg of neutralizing antibody against IFN-γ (Leinco Technologies, Inc., I-1190) (n = 14) 12 h before i.p. injection of TNF-α and IFN-γ. Mice were monitored for survival. The results were pooled from 2 independent experiments.
5
Determination of Blood Counts and Serum Levels
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Blood was drawn by retro-orbital or submandibular bleeding (serial bleed, 35 μL) for determination of complete blood counts. The method of collection was kept consistent within the same time-course experiment. Serum was obtained from blood drawn by intracardiac bleeding. Serum erythropoietin (R&D MEP00B) and Ftl (Abcam, ab157713) levels were determined by enzyme-linked immunosorbent assays. Tissue non-heme iron concentrations were determined as described previously.15 Serum iron levels were determined using a kit (Fisher:23666320).
6
Lipid and Ferritin Measurement Protocol
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Triglyceride (TG) and total cholesterol (TCHO) levels in tissues were measured using ASAN SET TG-S and ASAN SET Total Cholesterol (ASAN PHARM., Seoul, Korea), respectively, according to the manufacturer’s protocol. Free fatty acid (FFA) was analyzed using an EnzyChrom Free Fatty Acid Assay Kit (Bio-Assay Systems, Hayward, CA, USA) according to the manufacturer’s instructions. Plasma was measured using a FUJIFILM DRI-CHEM 4000i machine (FujiFilm Co., Tokyo, Japan), and FUJI DRI-CHEM slides were used for TG, TCHO, and HDL-C (high density lipoprotein-cholesterol) (Fujifilm Co., Tokyo, Japan). LDL-C (low density lipoprotein-cholesterol) levels were calculated using Frieldwann’s equation: LDL-C = [TCHO − (HDL-C + (TG/5))]. Plasma ferritin was assessed by using a commercially available enzyme linked immunosorbent assay (ELISA) kit (ab157713, Abcam, Cambridge, MA, USA).
7
Cytokine-Induced Mortality in Mice
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Age- and gender-matched 6- to 8-week-old mice of the indicated genotypes were injected intraperitoneally with 10 μg TNF-α alone, 20 μg IFN-γ alone, or TNF-α + IFN-γ diluted in DPBS. Animals were under permanent observation, and survival was assessed every 30 min. Blood was collected 5 h after cytokine injection. Blood composition was analyzed using an automated hematology analyzer. Serum LDH, ALT, AST, blood urea nitrogen (BUN), and ferritin were analyzed by colorimetry using respective kits (LDH, REF#A11A01824; ALT, REF#A11A01627; AST, REF#A11A01629; and BUN, REF#A11A01641, all from HORIBA; and ferritin, ab157713, Abcam) according to the manufacturer’s instructions.
For neutralizing antibody treatment, age- and gender-matched 6- to 8-week-old WT mice were administered intraperitoneally 200 μl of DPBS containing 500 μg of isotype control (Leinco Technologies, Inc., I-536) (n = 10) or 500 μg of neutralizing antibody against TNF-α (Leinco Technologies, Inc., T-703) plus 500 μg of neutralizing antibody against IFN-γ (Leinco Technologies, Inc., I-1190) (n = 14) 12 h before i.p. injection of TNF-α and IFN-γ. Mice were monitored for survival. The results were pooled from 2 independent experiments.
For neutralizing antibody treatment, age- and gender-matched 6- to 8-week-old WT mice were administered intraperitoneally 200 μl of DPBS containing 500 μg of isotype control (Leinco Technologies, Inc., I-536) (n = 10) or 500 μg of neutralizing antibody against TNF-α (Leinco Technologies, Inc., T-703) plus 500 μg of neutralizing antibody against IFN-γ (Leinco Technologies, Inc., I-1190) (n = 14) 12 h before i.p. injection of TNF-α and IFN-γ. Mice were monitored for survival. The results were pooled from 2 independent experiments.
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