The levels of TNF-α and IL-6 in the colon mucosa were measured using commercial mouse TNF-α and IL-6 ELISA kits (eBioscience, USA), according to the manufacturer’s instructions. The absorbance of the final products was measured on a microplate reader at a wavelength of 450 nm. The results were expressed as pg/mg tissue. Eight mice were randomly selected from each group for ELISA assays.
Mouse tnf α and il 6 elisa kits
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Mouse TNF-α and IL-6 ELISA kits are laboratory assays used to quantitatively measure the levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in mouse biological samples. These kits provide a quantitative, sandwich enzyme-linked immunosorbent assay (ELISA) method for the in vitro determination of these analytes.
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Lab products found in correlation
Anti ly6g, by Abcam (1 mentions)
Anti f4 80, by Santa Cruz Biotechnology (1 mentions)
P c jun, by Cell Signaling Technology (1 mentions)
C jun, by Cell Signaling Technology (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
4 protocols using mouse tnf α and il 6 elisa kits
1
Quantifying Inflammatory Cytokines in Colon Mucosa
2
Quantifying TNF and IL-6 Cytokines
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For the measurement of tumor necrosis factor (TNF) and interleukin-6 (IL-6) production, supernatants from cell cultures stimulated as indicated were harvested and cytokines were quantified using mouse TNFα and IL-6 ELISA kits (eBioscience), according to the manufacturer’s instructions.
3
Evaluation of Compound 34# in LPS-Induced Inflammation
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Lipopolysaccharide (LPS) was obtained from Sigma-Aldrich (St.Louis, MO, USA). Antibodies for P38, p-P38, JNK, p-JNK, ERK, p-ERK, p-c-jun, c-jun, GAPDH, and IκB were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-F4/80 was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-LY6G was obtained from Abcam (Cambridge, UK). Antibodies for CD11B, CD45 and LY6Cwere obtained from BD Pharmingen (New York, USA). The mouse TNF-α and IL-6 ELISA kits were purchased from eBioscience (San Diego, CA, USA). Compound 34# was synthesized in our lab and structurally identified using MS and 1 H-NMR analyses. For in vivo studies, 34# was dissolved in 0.5% carboxymethylcellulose sodium (CMCNa) in water solution and 0.5% CMCNa alone was used as vehicle control. For cell culture studies, 34# was dissolved in dimethyl sulfoxide (DMSO), and the same volume of DMSO alone was used as the vehicle control.
4
Colorectal Tumor Load and Cytokine Analysis
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After 12 weeks, the mice were anesthetized with diethyl ether and sacrificed via transcardiac perfusion and colon tissues were removed. The colons were dissected longitudinally along the main axis and washed with 0.9% saline. The diameter of each tumor was measured using a sliding caliper, and then total tumor load of each colon was calculated as the sum of the diameter of each tumor. The process of measurement and calculation was blinded by the operator. Subsequently, the whole colon was divided into sections for ELISA and histopathological examination. The remaining colon was stored at -80 °C.
2.1.4. ELISA for tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-10 in colon mucosa
The levels of TNF-α and IL-6 in the colon mucosa were measured using commercial Mouse TNF-α and IL-6 ELISA Kits (eBioscience). The absorbance of the final colored product was measured at 450 nm as the primary wavelength. The results were expressed as pg/mg. Eight mice were randomly selected from each group for ELISA.
2.1.4. ELISA for tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-10 in colon mucosa
The levels of TNF-α and IL-6 in the colon mucosa were measured using commercial Mouse TNF-α and IL-6 ELISA Kits (eBioscience). The absorbance of the final colored product was measured at 450 nm as the primary wavelength. The results were expressed as pg/mg. Eight mice were randomly selected from each group for ELISA.
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