Ab255729

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab255729 is a primary antibody product offered by Abcam. It is a polyclonal antibody raised against a specific target. The core function of this product is to specifically bind and detect the target antigen.

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Lab products found in correlation

13 protocols using ab255729

Quantification of Inflammation-Related Cytokines by ELISA

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Using enzyme linked immunosorbent assay (ELISA), inflammation-related cytokines including IL-1β, TNFα, IL-6, IL-12, IL-4, and IL-10 were detected and quantified 24 h after ICH [34 ]. The manufacturer’s protocol was followed as provided in the ELISA kit. In brief, brain tissue extracted, and protein supernatant was acquired as described above. The protein sample or standard was mixed with antibody cocktail in pre-coated 96-well microplate and then incubated at 25 °C for 1 h. Next, the wells were washed using wash buffer three times. The development solution was added to each well to co-incubate in a microplate, and stop solution was added, and the microplate was analyzed using a spectrophotometer to calculate optical density (OD) at 450 nm. By comparing the OD of target protein with standard curve, the protein concentration was quantified. The following ELISA kits were used: IL-1β (ab197742, Abcam, MA, USA), TNFα (ab208348, Abcam, MA, USA), IL-6 (ab222503, Abcam, MA, USA), IL-12 (ab236717, Abcam, MA, USA), IL-4 (ab221833, Abcam, MA, USA), and IL-10 (ab255729, Abcam, MA, USA).

Li T., Xu W., Ouyang J., Lu X., Sherchan P., Lenahan C., Irio G., Zhang J.H., Zhao J., Zhang Y, & Tang J. (2020). Orexin A alleviates neuroinflammation via OXR2/CaMKKβ/AMPK signaling pathway after ICH in mice. Journal of Neuroinflammation, 17, 187.

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Cytokine and Biomarker Quantification

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Sera and lysate from in vivo models (n = 7–9 mice per treatment group) and cell culture media supernatants from treated PM_ϕ were used to assess the concentrations of proinflammatory cytokines (IL-1β, IL-6, and TNFα), anti-inflammatory cytokines (IL-10 and TGF-β), troponin I and cAMP. IL-1β (ab197742; Abcam), IL-6 (ab222503; Abcam), TNFα (ab208348; Abcam), IL-10 (ab255729; Abcam), TGF-β (KE10005; Proteintech), cAMP (JL13362; Jianglai Bio, China), and troponin I (JL31923; Jianglai Bio, China) ELISA were performed in triplicates as per the instructions of the manufacturer.

Adzika G.K., Hou H., Adekunle A.O., Rizvi R., Adzraku S.Y., Li K., Deng Q.M., Mprah R., Ndzie Noah M.L., Adu-Amankwaah J., Machuki J.O., Shang W., Ma T., Koda S., Ma X, & Sun H. (2021). Amlexanox and Forskolin Prevents Isoproterenol-Induced Cardiomyopathy by Subduing Cardiomyocyte Hypertrophy and Maladaptive Inflammatory Responses. Frontiers in Cell and Developmental Biology, 9, 719351.

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Quantifying Mouse Serum Cytokine Levels

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To measure blood cytokine levels, serum samples were obtained by coagulating whole blood in EDTA tubes for 10 min at room temperature (i.e., 25 ± 1°C) and centrifugation for 10 min at 2,000 × g (n=6 per group). ELISA kits for mouse IL-10 (ab255729, Abcam Plc, Cambridge, UK) and mouse IL-1 beta (ab197742, Abcam Plc, Cambridge, UK) were used to detect inflammation levels performed according to the manufacturer’s instructions. Each sample was examined at least twice and analyzed using the ELISAcalc software (V0.1, Blue Gene).

Ding X., Zhou J., Zhao L., Chen M., Wang S., Zhang M., Zhang X, & Jiang G. (2022). Intestinal Flora Composition Determines Microglia Activation and Improves Epileptic Episode Progress. Frontiers in Cellular and Infection Microbiology, 12, 835217.

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Serum Cytokine Analysis by ELISA

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The levels of midkine, IL-10, and IFN-γ in serum samples or conditional medium were evaluated by ELISA kit (Abcam # ab193761, # ab279416, # ab185986, #ab255729 and #ab174443, USA) according to manufacturer’s instructions. Briefly, samples were incubated with antibody-coated plates for 1 h at room temperature and washed for five times. Then plates were incubated with biotinylated antibody, streptavidin antibody, substrate solution, and stop solution consecutively. The absorbance at 450 nm was measured by a microplate reader. All samples were done in triplicates.

Ding L., Wang N., Wang Q., Fan X., Xin Y, & Wang S. (2023). Midkine inhibition enhances anti-PD-1 immunotherapy in sorafenib-treated hepatocellular carcinoma via preventing immunosuppressive MDSCs infiltration. Cell Death Discovery, 9, 92.

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Quantifying Cytokine Levels Using ELISA

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The serum concentrations of IFN-γ, IL-6, IL-10, and IL-22 were measured using the following ELISA kits according to the manufacturer’s protocols: IFN-γ (Catalog No. ab100689, Abcam), IL-6 (Catalog No. ab222503, Abcam), IL-10 (Catalog No. ab255729, Abcam), and IL-22 (Catalog No. ab223857, Abcam). Briefly, standards, blank controls, and test samples (100 μL/well) were added to 96-well plates. Next, a biotinylated antibody (50 μL/well) was added to each well and incubated at 37 °C for 90 min. Unbound biotinylated antibody was removed using washing buffer (PBS with a mild detergent). Streptavidin–horseradish peroxidase (100 μL/well) was added to the plates, followed by incubation at 37 °C for 30 min. TMB (100 μL/well) was added, followed by incubation at 37 °C for 10 min in the dark. The reaction was interrupted using a stop solution, and optical density was measured at a wavelength of 450 nm using a microplate reader. The concentration of each sample was quantified using a standard curve.

Li K., Cui Y., Zheng X., Min C., Zhang J., Yan Z., Ji Y., Ge F., Ji H, & Zhu F. (2024). Jian Gan powder ameliorates immunological liver injury in mice by modulating the gut microbiota and metabolic profiles. European Journal of Medical Research, 29, 240.

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Evaluating Anti-Inflammatory Effects of HGs

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To assess the anti-inflammatory effect of 10HA and 10HA 120NP HGs further, ELISA assays were performed to quantify the release of six pro-inflammatory mediators by LPS-stimulated RAW264.7. Mouse TNF-α, IL-6, IL-10, IL-12p40, IL-23, and PGE2 ELISA kits were purchased from abcam (ab208348, ab222503, ab255729, ab236717, ab119545, and ab133021, Cambridge UK). In brief, cells were seeded on top of the HGs and CGs at a density of 2 × 10⁶ cells/well and cultured for 24 h. Then, cells were exposed to medium containing LPS (500 ng/mL) and incubated overnight. Media were collected and stored at −20 °C until use. Levels of mouse TNF-α, IL-6, IL-10, IL-12p40, IL-23, and PGE2 in cell culture supernatants were determined by the corresponding ELISA kit according to the protocol recommended by the manufacturer. Experiments were performed using six replicates per formulation. ANOVA was performed at significance levels of p < 0.05, p < 0.01, and p < 0.001.

Halfter N., Espinosa-Cano E., Pontes-Quero G.M., Ramírez-Jiménez R.A., Heinemann C., Möller S., Schnabelrauch M., Wiesmann H.P., Hintze V, & Aguilar M.R. (2023). Ketoprofen-Based Polymer-Drug Nanoparticles Provide Anti-Inflammatory Properties to HA/Collagen Hydrogels. Journal of Functional Biomaterials, 14(3), 160.

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Cytokine Quantification by ELISA

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Cytokines were determined in the culture medium by enzyme-linked immunoassay (ELISA) using the ultrasensitive ELISA kits for IL-1β (Abcam, ab197742), IL-4 (Abcam, ab100710), IL-6 (Abcam, ab222503), IL-10 (Abcam, ab255729), TGFβ (Abcam, ab119557) and TNF-α (Abcam, ab208348) according to the manufacturer’s instructions. Briefly, the samples were obtained from the culture medium after treatments. The medium was centrifuged at 2,000 rpm to remove cell debris and diluted 50% with distilled water. The concentration of cytokines was determined based on the standard curve provided by the manufacturer and expressed as pg/ml.

Hernandez-Espinosa D.R., Gale J.R., Scrabis M.G, & Aizenman E. (2023). Microglial reprogramming by Hv1 antagonism protects neurons from inflammatory and glutamate toxicity. Journal of neurochemistry, 165(1), 29-54.

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Quantifying Cytokine Levels in Blood and BALF

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The concentrations of IL-18 (ab215539, abcam), IL-1β (ab217608, abcam), IL-17A (ab216167, abcam) and IL-10 (ab185986, abcam) in human peripheral blood serum, and IL-4 (ab100710, abcam), IL-5 (ab204523, abcam), IL-18 (ab216165, abcam), IL-1β (ab241673, abcam), IL-17A (ab199081, abcam) and IL-10 (ab255729, abcam) in mouse BALF were determined by ELISA.

Wu J., Wang P., Xie X., Yang X., Tang S., Zhao J., Liu T., Wang J., Zhang J., Xia T, & Feng X. (2024). Gasdermin D silencing alleviates airway inflammation and remodeling in an ovalbumin-induced asthmatic mouse model. Cell death & disease, 15(6).

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Cytokine Quantification via ELISA

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Under the instructions of the producer, levels of interleukin (IL)-10 (ab255729, Abcam), IL-6 (ab222503, Abcam), and IL-1β (ab197742, Abcam) were determined using ELISA kits. The absorbance was determined using a microscope reader (Bio-Rad) [24 (link)]. Each experiment was performed in triplicate.

Chen Z., Zheng B., Zhang Z, & Huang Z. (2023). Protective role of FBXL19 in Streptococcus pneumoniae-induced lung injury in pneumonia immature mice. Journal of Cardiothoracic Surgery, 18, 92.

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Measuring Nutritional and Inflammatory Markers

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Serum samples were collected at week 12, and biochemical nutritional markers TC, TG, HDL, alanine transaminase (ALT), aspartate transaminase (AST), and inflammatory cytokines (IL-1β and IL-10) were measured. Biochemical, and nutritional markers were measured using a Fuji DRI-CHEM Clinical Chemistry Analyzer FDC 3500 (Fujifilm, Tokyo, Japan), and LDL level was calculated using the Friedewald formula^{42 (link)}. Serum inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) kits (IL-1β, Abcam, ab197742; IL-10, Abcam, ab255729). Experiments were performed in accordance with the manufacturer’s instructions. Optical densities were measured at 450 nm using a spectrophotometer (SpectraMax ABS Plus, San Jose, CA, USA).

Lee D.J., Kang A.N., Lee J., Kwak M.J., Mun D., Lee D., Oh S, & Kim Y. (2024). Molecular characterization of Fusarium venenatum-based microbial protein in animal models of obesity using multi-omics analysis. Communications Biology, 7, 133.

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