Wells were coated overnight at 4°C with anti-mouse IL-10 (R and D Systems) in coating buffer (0.0125M sodium bicarbonate plus 0.0875 M anhydrous sodium carbonate, pH 9.6). Wells were washed three times each with 0.05% Tween-20 in PBS (PBST, pH 7.4) and deionized water (DW), then blocked with 3% bovine serum albumin (BSA) in PBS for 1 h at room temperature (RT). After washing, samples and standards were added and incubated for 1 h at RT. Again, after washing, subsequent steps were performed with biotin-conjugated anti-mouse IL-10 followed by avidin-conjugated with horseradish peroxidase (HRP-Avidin; BioLegend) and 3,3′,5,5′-tetramethylbenzidine (TMB) substrate (BioLegend), all with wash steps in between. IL-10 was detected following the addition of 100 μl of 2 N sulfuric acid (H2SO4) to terminate the reaction. Optical density (OD) was measured at 450 nm within 30 min of acid addition. Quantification of IL-10 was performed by generation of a standard curve using recombinant mouse IL-10.
Anti mouse il 10
Manufactured by R&D Systems
Sourced in United States
Anti-mouse IL-10 is a mouse interleukin-10 (IL-10) antibody. IL-10 is an anti-inflammatory cytokine that plays a role in regulating the immune response. The Anti-mouse IL-10 antibody can be used for the detection and quantification of mouse IL-10 in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Spectramax m5 microplate reader, by Molecular Devices (2 mentions)
Mouse anti il12, by BD (2 mentions)
Phosphatase conjugated streptavidin, by Southern Biotech (2 mentions)
P nitrophenyl phosphatase pnpp substrate, by Southern Biotech (2 mentions)
Softmax pro acquisition and analysis software, by Molecular Devices (2 mentions)
Avidin hrp, by BioLegend (1 mentions)
3 3 5 5 tetramethylbenzidine tmb substrate, by BioLegend (1 mentions)
Normal goat igg control, by R&D Systems (1 mentions)
Anti mouse vegf, by R&D Systems (1 mentions)
Anti human vegf, by R&D Systems (1 mentions)
Ct dna, by Merck Group (1 mentions)
4 protocols using anti mouse il 10
1
Quantitative Mouse IL-10 ELISA Protocol
2
Blocking Antibodies and Mcl-1 Inhibitor Treatment
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The following blocking antibodies were used: 10–20 μg/mL anti-mouse GM-CSF blocking antibody (eBioscience, San Diego, CA, USA), 20 μg/mL anti-mouse VEGF, 20 μg/mL anti-mouse IL-10, 10 μg/mL anti-human GM-CSF, 10 μg/mL anti-human VEGF and 20 μg/mL anti-human IL-10 blocking antibodies (R&D systems, Minneapolis, MN, USA). Isotype controls used were 10 μg/mL rat IgG2aK isotype control (eBioscience) and normal goat IgG control (R&D). Mcl-1 inhibitor MIM1 (R&D) was dissolved in DMSO.
3
Quantifying Cytokine Levels via ELISA
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ELISA was performed based on the protocol described previously10 (link). Immunol 2HB-microtiter plates were coated with mouse anti IL-10 (R&D Systems) or mouse anti-IL12 (BD Biosciences) followed by blocking with 2% BSA-PBS for 2 hr. Culture supernatants or diluted sera were added after washing and incubated overnight at 4 °C. Secondary Ab labeled with biotin were added to plates, and incubated for 2 hr. Next plates were developed by phosphatase-conjugated streptavidin (AKP) followed by the addition of p-nitrophenyl phosphatase (pNPP) substrate (Southern Biotech). Optical density was measured using a SpectraMax M5 microplate Reader and SoftMax Pro Acquisition and Analysis software (Molecular Devices).
4
Quantification of Anti-Cytokine Antibodies
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