Müller cells harvested from σR1+/+ and σR1−/− mice were incubated in serum-free media in presence/absence of 1.0 µg/ml LPS for 24 h. Conditioned media was used in ELISA assays to detect secretion of MIP1γ, MIP2, MIP3α and IL12 (p40/p70). ELISA assays were performed using RayBiotech Mouse ELISA kits: ELM-MIP1γ, ELM-MIP2, ELM-MIP3α and ELM-IL12 (p40/p70) following the manufacturer’s protocol. The 96-well plate (provided with the kit) is coated with immobilized antibody. After the sample was applied to the coated plate, the specific biotinylated antibody (provided with the kit) was added. The HRP-conjugated streptavidin was added followed by tetramethylbenzidine to yield a color reaction, which was detected at an absorbance of 450 nm using the VERSAmax Tunable Microplate Reader (Molecular Devices, Sunnyvale, CA).
Mouse elisa kit
Manufactured by RayBiotech
Sourced in United States
The Mouse ELISA kit is a laboratory equipment product designed for the quantitative measurement of specific target analytes in mouse samples. It utilizes the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the presence of the target analyte in the sample.
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Lab products found in correlation
9 protocols using mouse elisa kit
1
Cytokine Secretion in Müller Cells
2
Quantifying Cytokine Levels in Mouse Joints
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The mouse joints were pulverised using a mortar and pestle after freezing in liquid nitrogen. Tissue was transferred to 15‐mL tubes, placed on dry ice and resuspended in 1 mL PBS per 200 mg of tissue (containing Complete™ Protease Inhibitor Cocktail) and homogenised using a homogeniser. Mouse joint homogenates were centrifuged for 10 min at 500 g at 4°C. Supernatants were transferred to Eppendorf tubes and centrifuged at 15000 g for 5 min and collected for cytokine analysis. Concentrations of IFN‐γ, IL‐6 and TNF‐α in joint lysates or plasma were measured using mouse ELISA kits from RayBiotech (Peachtree Corners, USA, Cat. #ELM‐IFNg‐1, ELM‐IL‐6‐1 and ELM‐TNFa‐1), following the manufacturer's instructions. Briefly, 100 μL standards or samples was pipetted into the wells and incubated for 2.5 h at room temperature to allow the cytokine in a sample to be bound to the wells by the immobilised antibody. The wells were washed three times, and 100 μL biotinylated anti‐Mouse cytokine antibody was added. After washing away unbound biotinylated antibody, 100 μL HRP‐conjugated streptavidin was added to the wells. The wells were again washed three times, and a 100 μL TMB substrate solution was added to the wells. A volume of 50 μL Stop Solution was added to the wells, and the OD was measured at 450 nm.
3
Hippocampal Inflammatory Markers and Amyloid-β
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Hippocampal TNF-α, IL-6 levels, NF-κB p65, and β-amyloid1-42 were measured using mouse ELISA kits purchased from RayBiotech Inc. (Norcross, GA, USA) and R&D Systems Inc. (Minneapolis, USA), respectively. The procedures were performed according to the manufacturers' instructions. Results are presented as pg/mg protein for TNF-α, IL-6, NF-κB p65, and β-amyloid1-42.
4
Hippocampal Protein Markers Quantification
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Hippocampal IL-6 and ICAM-1 levels were estimated using mouse ELISA kits purchased from RayBiotech Inc. (Norcross, Georgia, USA) and MyBioSource Inc. (San Diego, CA, USA), respectively. Cleaved caspase-3 and amyloid-β1-42 mouse ELISA kits were provided from Cusabio, Wuhan, China. The hippocampal levels of these markers were measured according to the manufacturer's instructions for each respective ELISA kit and expressed as their corresponding units to the tissue protein content determined by Salama et al. [44 (link)]. The ELISA assay measures the amount of sample by sandwiching it between two antibodies, one of which is precoated to the microtiter plate, and the other of which acts as a detector antibody. The microtiter plate provided in each kit was precoated with an antibody specific to each marker. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for the marker, and avidin conjugated to HRP is added to each microplate well and incubated. Then, a TMB substrate solution is added to each well. Only those wells that contain the protein, biotin-conjugated antibody, and enzyme-conjugated avidin will exhibit a change in color. The enzyme–substrate reaction is terminated by the addition of a sulfuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450 nm.
5
TNFα Expression in Cell Hydrogels
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Raw 264.7 cells were encapsulated in control and PEG-DGEA hydrogels to assess differences in TNFα expression due to immobilized DGEA. Post-treatment with M1 media, conditioned media, or cell supernatant was collected. The concentrations of TNFα were measured by utilizing a mouse ELISA kit (RayBiotech) and following manufacturer’s instructions. Student's t test was used to determine differences in cytokine expression between the treatment groups.
6
Mouse Cytokine Profiling Protocol
7
Measuring Tissue SDF-1α Levels
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SDF-1α is an important isoform of SDF-1. To inspect the changes in SDF-1α, the LVs were isolated and maintained at -80°C until use. The tissue SDF-1α was measured with a mouse ELISA kit (RayBiotech Inc., Norcross, GA, USA) according to the manufacturer’s instructions as described previously [15 (link)].
8
Inflammatory Cytokine Quantification
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Blood was obtained and centrifuged at a speed of 3000 rpm for 10 min at 4°C. Serum was collected for assessing the levels of inflammatory cytokines TGF-β and IL-17 by Mouse ELISA Kit (RayBiotech, USA). IL-1β, IL-6, and TNF-α were quantified in lung homogenates using commercial ELISA kits (eBiosciences).
9
Protein and Cytokine Analysis in BALF
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The total protein concentration in the BALF was measured by using a BCA Protein Assay kit (Biomiga, USA) according to the manufacturer's instructions. Tumour necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the BALF were analyzed by using a mouse ELISA kit (RayBiotech, USA) according to the manufacturer's instructions. BALF level of high-mobility group box 1 protein (HMGB1) were detected by ELISA kit (Cusabio, China) according to the manufacturer's protocol.
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