IFNγ, IL-4, IL-5, IL-6, IL-10 and IL-12p40/23 levels on culture supernatants were quantified by interleukin-specific sandwich ELISA assays (BD Bioscience, NJ, USA). MIP-1α and MIP-2 were also detected in the culture media using sandwich ELISA assays, according to the instructions of the manufacturer (RayBiotech, Inc., GA, USA). In some cases, cytokines were detected by qRT-PCR using a Corbett Rotor Gene 6000 Real-Time PCR Machine and the SYBR Green 1 dye (Applied Biosystem). Standard amplification conditions were 3 min at 95°C and 40 cycles of 10 s at 95°C, 30 s at 60°C, and 30 s at 72°C. For detection of cytokines the following primers were used: IFNγ: F: 5′-GGAGGAACTGGCAAAAGGATGGTGA-3′ and R: 5′-GCGCTGGACCT-GTGGGTTGT-3′; IL-4: F: 5′-AGGTCACAGGAGAAGGGACGCC-3′ and R: 5′-TGC-GAAGCACCTTGGAAGCCC-3′; IL-10: F: 5′-TTCCCAGTCGGCCAGAGCCA and R: 5′-GGGGAGAAATCGATGACAGCGCC-3′. Results were expressed as the ratio between each evaluated cytokine and GAPDH expression. For GAPDH detection, sense and antisense primers were 5′-TCGGAGTCAACGGATTG-3′ and 5′-CCTGGAAGAT-GGTGATGG-3′, respectively.
Interleukin specific sandwich elisa assays
Manufactured by BD
Sourced in United States
Interleukin-specific sandwich ELISA assays are laboratory equipment used to detect and quantify specific interleukin proteins in biological samples. These assays employ a sandwich-based enzyme-linked immunosorbent assay (ELISA) technique to measure the concentration of target interleukins.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using interleukin specific sandwich elisa assays
1
Quantification of Cytokine Levels
2
Dendritic Cell Maturation Assay with Helminth Antigens
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone Marrow-derived Dendritic Cells (BMDCs) were generated from bone marrow precursors from BALB/c mice. Briefly, bone marrow precursor cells were harvested and plated at a density of 2–5 × 105 cells/ml in complete culture medium supplemented with GM-CSF-containing supernatant. After 3 days of culture at 37°C, the medium was replaced. Cells were recovered on day 8 and analyzed for the expression of CD11c by flow cytometry. To analyze DC-maturation, BMDCs (2.5 × 105/well) were incubated at 37°C and 5% CO2 in 96-well plates with FhTE, FhPox, FhCB (75 μg/ml) or medium alone in the presence or absence of LPS (Escherichia coli 0111:B4, 0.5–1 μg/ml) overnight at 37°C. Alternatively, cells were pre incubated for 45 min. at 37°C with 10 mM of monosaccharides (Man, GalNAc or arabinose) or 10 μM of specific signaling inhibitors (PHPS1; GW5074; and ER27319). Cells were then centrifuged at 1,500 rpm for 5 min at 4°C and supernatants were then collected. Cytokine (IL-12/23p40, IL-10 and IL-6) levels were tested on culture supernatants by interleukin specific sandwich ELISA assays (BD Bioscience, NJ, USA).
3
Evaluating Immune Responses to Fasciola hepatica
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A group of five animals were orally infected with ten F. hepatica metacercariae (Baldwin Aquatics, OR, USA) per animal. Following 3 weeks of infection, mice were bled and necropsied, and the livers, spleens, hepatic-draining lymph nodes (HLNs) and peritoneal exudates cells (PECs) were removed. Splenocytes or PECs (2.5-5×10 6 cells/mL) were cultured in complete medium consisting of RPMI-1640 with glutamine supplemented with 10 % heat-inactivated foetal bovine serum (FBS), 50 μM 2-mercaptoethanol, 100 U/mL penicillin, and 100 mg/mL streptomycin for 72 h in the presence or absence of Fhmuc peptide or FhESP (10 μg/mL). Secreted cytokine (IFNγ, IL-5, and IL-17) levels of culture supernatants were measured by interleukin-specific sandwich ELISA assays (BD Biosciences, NJ, USA). Antibody reactivity against Fhmuc or FhESP from obtained sera was analysed by ELISA. Then, hepatic damage was histologically analysed, confirming >80 % tissue damage in infected animals. Naive animals were used as control group (n=5).
4
Intraperitoneal Immunization and Immune Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice (n=8 per group) were immunised intraperitoneally (i.p.) with Fhmuc (20 μg) or PBS (control group) in complete Freund's adjuvant on day 0 followed by two additional injections in incomplete Freund's adjuvant on days 14 and 28. Two weeks after the final immunisation, mice were sacrificed and the spleens, HLN, and PECs were removed. PECs were harvested by washing the peritoneal cavity with 10 mL of cold PBS. Cells were dispersed manually, centrifuged at 1000g for 5 min, and suspended (1×10 6 cells/well) in complete culture medium. Cells were incubated in 96-well plates with Fhmuc peptide or FhESP (10 μg/mL) or medium alone for 72 h at 37 °C with 5 % CO 2 . Secreted cytokine (IFNγ, IL-5, and IL-17) levels of culture supernatants were measured by interleukin-specific sandwich ELISA assays (BD Biosciences, NJ, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!