Anti–mouse IFN-β neutralizing antibody (300 ng/mouse; PBL Assay Science, 32400-1), rabbit polyclonal IgG control (300 ng/mouse; BioLegend, CTL-4112), anti-mouse monoclonal IFNAR1 neutralizing antibody (MAR1; 1 μg/mouse; Leinco Technologies, I-401), and mouse IgG control (1 μg/mouse; Leinco Technologies, I-536) were dissolved in sterile PBS and administered intrathecally as described previously.
I 536
Manufactured by Leinco Technologies
The I-536 is a compact and versatile laboratory instrument designed for precise measurements. Its core function is to provide accurate and reliable data for various scientific applications.
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5 protocols using i 536
1
Intrathecal Antibody Administration
2
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.
3
Therapeutic Targeting of Cytokines in SARS-CoV-2 Infection
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Age- and gender-matched, 7- to 8-week old K18-ACE-2 transgenic mice were anesthetized with 5% isoflurane and then infected intranasally with SARS-CoV-2 in 50 μl DPBS containing around 2 × 104 PFU. Infected mice were administered intraperitoneally 200 μl of DPBS containing 500 μg of isotype control (Leinco Technologies, Inc., I-536) (n = 12) 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 = 13) on days 1, 3, and 4 post-infection. Mice were monitored over a period of 14 days for survival.
4
SARS-CoV-2 Infection in Transgenic Mice
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Age- and gender-matched, 7- to 8-week old K18-ACE-2 transgenic mice were anesthetized with 5% isoflurane and then infected intranasally with SARS-CoV-2 in 50 μL DPBS containing around 2 × 104 PFU. Infected mice were administered intraperitoneally 200 μL of DPBS containing 500 μg of isotype control (Leinco Technologies, Inc., I-536) (n = 12) 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 = 13) on days 1, 3, and 4 post-infection. Mice were monitored over a period of 14 days for survival.
5
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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