Ab119558

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab119558 is a rabbit polyclonal antibody that targets the human Histone H3 protein. The antibody is suitable for use in various applications, including Western blotting, immunohistochemistry, and immunocytochemistry.

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Lab products found in correlation

9 protocols using ab119558

Quantifying TGF-β1 in Heart Tissue

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First LV heart tissues were removed and placed in ice-cold saline. Afterward, tissues were homogenized and centrifuged for 10 minutes at 1000 rpm, and finally, the supernatant was separated. Then, the level of TGF-β1 in LV heart tissues was measured using the ELISA method according to the kit instructions (Cat.No: ab119558, Abcam, MA, USA) [36 (link)].

Hajializadeh Z., Khaksari M., Dabiri S., Darvishzadeh Mahani F., Raji-Amirhasani A, & Bejeshk M.A. (2023). Protective effects of calorie restriction and 17-β estradiol on cardiac hypertrophy in ovariectomized obese rats. PLOS ONE, 18(4), e0282089.

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Cytokine Levels in BALF

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The levels of proinflammatory cytokines (TGF-β, IL-1β, IL-6, and TNF-α) in BALF were performed according to the instruction of ELISA kits (Abcam: ab119558, ab100705, ab178013 and ab181421) (Abcam, Cambridge, MA, USA). The sensitivities of ab119558, ab100705, ab178013 and ab181421 ELSIA kits were 8, 5, 5 and 14 pg/mL, respectively.

Huang X., Zhu J., Jiang Y., Xu C., Lv Q., Yu D., Shi K., Ruan Z, & Wang Y. (2019). SU5416 attenuated lipopolysaccharide-induced acute lung injury in mice by modulating properties of vascular endothelial cells. Drug Design, Development and Therapy, 13, 1763-1772.

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Evaluating Hepatoprotective Potential

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Carbon tetrachloride (CCl₄) (#PHR1063), silymarin (#S0292), and other chemicals and reagents were purchased from Sigma Aldrich Chemicals Co., St. Louis, Missouri, United States. Kits for the pro-inflammatory (TGF-β #ab119558, TNF-α #ab236712, IL-6 #ab234570, and adiponectin #ab239421) and anti-inflammatory cytokine markers (SOD #ab65354, CAT #ab118184, GSR #ab65322, and TBARS/MDA #ab238537), liver function test (LFT) (AST #ab263882, ALT #234579, ALP #ab83369, and Bil #ab235627) enzymes to detect free radical scavenging activity, mitochondrial membrane potential (MMP) (MMP #ab113852), and ROS (ab #ab186027) detection were purchased from Abcam, Cambridge, United Kingdom.

Alnuqaydan A.M., Almutary A.G., A. Alsahli M., Alnasser S, & Rah B. (2022). Tamarix articulata Induced Prevention of Hepatotoxicity Effects of In Vivo Carbon Tetrachloride by Modulating Pro-Inflammatory Serum and Antioxidant Enzymes to Reverse the Liver Fibrosis. Antioxidants, 11(9), 1824.

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Inflammatory Cytokine Analysis in Atrial Tissue

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Atrial tissue was minced and homogenized using a tissue homogenizer (TissueRuptor, QIAGEN) according to the manufacturer protocol. Inflammatory cytokines and chemokines were measured using a multiplex Luminex-based assay (LXSARM, R&D Systems). Each sample was run in duplicate in a 96-well plate. IL-1β, IL-2, IL-18, and TNF-α were measured using a MAGPIX system (C4447b, Luminex). Acquired mean fluorescence data were analyzed and calculated by xPONENT software. IL-6 (ERA32RB, Invitrogen), TGF-β1 (ab119558, Abcam), PDGF-AB (ab213906, Abcam), and MCP-1 (ab100778, Abcam) were quantified using commercially available ELISAs according to the manufacturers’ protocols.

Parent S., Vaka R., Risha Y., Ngo C., Kanda P., Nattel S., Khan S., Courtman D., Stewart D.J, & Davis D.R. (2023). Prevention of atrial fibrillation after open-chest surgery with extracellular vesicle therapy. JCI Insight, 8(15), e163297.

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Lung Cytokine Levels by ELISA

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Lung cytokine levels were analyzed using an enzyme-linked immunosorbent assay (ELISA). The levels of interleukin 1 beta (IL1-β; catalog no: 900-M91; Prepotech^®, USA), interleukin 6 (IL-6; catalog no: 900-M86; Prepotech^®, USA), interleukin 10 (IL-10; catalog no: ab100765; Abcam, UK), tumor necrosis factor-alpha (TNF-α; catalog no: ab100785; Abcam, UK) and transforming growth factor-beta (TGF-β; catalog no: ab119558; Abcam, UK) were determined according to the manufacturers’ protocols.

Seenak P., Kumphune S., Prasitsak T., Nernpermpisooth N, & Malakul W. (2022). Atorvastatin and ezetimibe protect against hypercholesterolemia-induced lung oxidative stress, inflammation, and fibrosis in rats. Frontiers in Medicine, 9, 1039707.

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Quantitative Cytokine and Inflammasome Profiling

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Kits purchased from Abcam, Waltham, USA (CAT # ab119558 and ab214566), were utilized for assessment of tissue TGF-β1 and IL10 levels, respectively. IFN-α levels were quantified using kits supplied by CUSABIO, Houston, TX 77054, USA (CAT #CSB-E08637r). Assay of NLRP3 levels was executed using kits provided by Aviva Systems Biology Co., San Diego, CA 92121, USA (CAT # OKCD04232). Determination of the fore-mentioned parameters was carried out following the vendor’s protocol.

Kabel A.M., Salama S.A., Adwas A.A, & Estfanous R.S. (2021). Targeting Oxidative Stress, NLRP3 Inflammasome, and Autophagy by Fraxetin to Combat Doxorubicin-Induced Cardiotoxicity. Pharmaceuticals, 14(11), 1188.

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Atrial Inflammation Biomarkers Analysis

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Homogenized atrial tissue obtained after the invasive electrophysiological study was analyzed for markers of atrial inflammation using ELISAs and a custom multiplex Luminex-based assay (LXSARM, R&D Systems). Interleukin 6 (IL-6, ERA32RB, Life Sciences), IL-17 (MBS2022678, MyBioSource), platelet-derived growth factor AB (PDGF-AB, ab213906, Abcam), monocyte chemoattractant protein-1 (MCP-1, ab100778, Abcam) and transforming growth factor beta 1 (TGFβ1; ab119558, Abcam) were analyzed using commercially available ELISAs. Four analytes (IL-1β, IL-10, IL-18, tumor necrosis factor alpha (TNFα)) were measured using a MAGPIX system (LXSARM-06) with post-hoc mean fluorescence quantification using xPONENT software. Atrial myeloperoxidase activity was evaluated using a colorimetric assay according to the manufacturer's directions (ab105136, Abcam).

Parent S., Vaka R., St Amant J., Kahn S., Van Remortel S., Bi C., Courtman D., Stewart D.J, & Davis D.R. (2024). Inactivation of the NLRP3 inflammasome mediates exosome-based prevention of atrial fibrillation. Theranostics, 14(2), 608-621.

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Tissue Homogenization and Protein Analysis

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Anastomotic tissue was homogenized in 1.5 mL PBS containing a proteinase inhibitor cocktail and 1 mM EDTA (pH 7.4) in tubes with stainless steel beads (Precellys® MK28 Lysing Kit). The tissue was disintegrated three times for 10 seconds at 6,500 rpm with 20-second breaks using the Precellys® 24 homogenizer. The homogenate was centrifuged at 20,000 x g for 30 minutes at 4°C, and the supernatant was stored at -80°C. MPO (EKR284; Nordic BioSite, Täby, Sweden) and TGF-β1 (ab119558; Abcam, Cambridge, UK) were analysed by rat ELISA kits. Soluble collagens were eluted from the tissue using a buffer (pH 6.0) containing 1 M NaCl for 24 hours at 4°C and determined by the Sircol assay [19 (link)]. MPO, TGF-β1, and soluble collagen were normalized to total protein determined by the Lowry assay.

Kjaer M., Russell W., Schjerling P., Cottarelli E., Christjansen K.N., Olsen D.M., Krarup P.M., Jessen L., Berner-Hansen M., Jorgensen L.N, & Ågren M.S. (2020). Glucagon-Like Peptide-2 Analogue ZP1849 Augments Colonic Anastomotic Wound Healing. Gastroenterology Research and Practice, 2020, 8460508.

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Quantifying Rat Inflammatory Proteins

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To examine the protein expression of TNF-α, TGF-β1 and ERK1/2, Rat TNF alpha ELISA (ab46070; Abcam, Cambridge, UK), Rat TGF-β1 ELISA (ab119558; Abcam) and ERK1 (pT202/pY204; Abcam) + ERK2 (pT185/pY187; Abcam) + total ERK1/2 ELISA (ab126445; Abcam) kits were used. According to the manufacturer's instructions, antibodies were first settled into 96-well plates and proteins from the RBMECs were then added. Next, biotinylated anti-TNF-α, biotin-conjugated anti-rat TGF-β1 monoclonal antibody, detection antibody Erk1 (T202/Y204)/ Erk2 (T185/Y187) and detection antibody Erk1/2 were mixed and incubated at room temperature. Thereafter, streptavidin-horseradish peroxidase (HRP) was pipetted into wells followed by washing with wash buffer. Next, TMB Substrate Reagent (BD Biosciences, Shanghai, China) was added and Stop Solution (BD Biosciences) was used for reaction termination. Color intensity was measured at 450 nm using a Varioskan ™ LUX Multimode Microplate Reader (Thermo Fisher Scientific). The results for this experiment were from 3 independent trials.

Chen W., Shen Z., Cai S., Chen L, & Wang D. (2021). FGF21 promotes wound healing of rat brain microvascular endothelial cells through facilitating TNF-α-mediated VEGFA and ERK1/2 signaling pathway. Advances in clinical and experimental medicine : official organ Wroclaw Medical University, 30(7).

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