Rabbit polyclonal anti tnfα antibody

Manufactured by Abcam

Sourced in United Kingdom

The Rabbit polyclonal anti-TNFα antibody is a laboratory reagent designed for the detection and research of tumor necrosis factor alpha (TNFα) in various biological samples.

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Rabbit anti- (5 citations)

3 protocols using rabbit polyclonal anti tnfα antibody

Quantifying Protein Expression in Ischemic Tissue

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Western blot analysis was used to detect protein expression in the ischemic tissue, as described by us in previous work^{21 (link),22 (link)}. Upon completion of electrophoresis, proteins were transferred to polyvinylidene fluoride membranes, which were incubated with primary antibodies (rabbit polyclonal anti-Bcl-2, rabbit polyclonal anti-BAX antibody, rabbit polyclonal anti-TN-Fα antibody, or rabbit polyclonal anti- IL-1β antibody; Abcam, Cambridge, MA, USA) for 24 h at 4°C. Next, membranes were washed with PBS three times for 6 min per wash, and then re-incubated with a secondary antibody (goat anti-rabbit IgG, Santa Cruz, Dallas, TX, USA; goat anti-mouse IgG, Abcam) for 1 h at room temperature. Finally, an enhanced chemiluminescence system was used to detect immunoreactive bands. Western blot images for each antibody, including β-actin, were analyzed using an image analysis program (Image J 1.42, NIH, Bethesda, MD, USA) to represent protein expression in terms of relative image density; mean protein expression from the control group was assigned a value of 1 to serve as a reference.

Li F., Yang Z., Stone C., Ding J.Y., Previch L., Shen J., Ji Y., Geng X, & Ding Y. (2019). Phenothiazines Enhance the Hypothermic Preservation of Liver Grafts: A Pilot in Vitro Study. Cell Transplantation, 28(3), 318-327.

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Western Blot Analysis of Neuroinflammatory Markers

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The ipsilateral hemispheres of the mouse brain samples were collected and homogenized in whole cell lysis buffer (KeyGEN BioTECH, China). The protein supernatants were centrifuged at 12,000 x g for 10 minutes at 4 °C and denatured at 100 °C. The proteins were subjected to SDS-PAGE (KeyGEN BioTECH, China) and transferred to PVDF membranes (Millipore, USA). Then, the membranes were blocked with 5% skim milk and incubated with a rabbit polyclonal anti-interleukin 1β (IL-1β) antibody (Abcam, UK), a rabbit polyclonal anti-TNFα antibody (Abcam, UK), a rabbit polyclonal anti-brain-derived neurotrophic factor (BDNF) antibody (Abcam, UK) and a rabbit anti-GAPDH antibody (Abcam, UK) overnight at 4 °C, followed by incubation with an HRP-linked secondary antibody (Cell Signaling Technology, USA) for 2 h at RT. Finally, the membranes were exposed and analyzed with ImageJ software.

Xu R., Bai Y., Min S., Xu X., Tang T, & Ju S. (2020). In vivo Monitoring and Assessment of Exogenous Mesenchymal Stem Cell-Derived Exosomes in Mice with Ischemic Stroke by Molecular Imaging. International Journal of Nanomedicine, 15, 9011-9023.

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Immunohistochemical Assessment of Hepatic Inflammation

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The sectioned slides were stained according to standard protocols described previously. Paraffin-embedded sections of hepatic tissue were deparaffinized, dehydrated, and stained immunohistochemically for detection of mouse monoclonal anti-CD68 antibody (diluted 1 : 200 in PBS; Abcam, UK), mouse monoclonal anti-TLR4 antibody (diluted 1 : 100 in PBS; Abcam, UK), rabbit polyclonal anti-MyD88 antibody (diluted 1 : 100 in PBS; Bioworld technology, Co. Ltd., Nanjing, China), rabbit polyclonal anti-NF-κBp65 antibody (diluted 1 : 1000 in PBS; Abcam, UK), rabbit polyclonal anti-IL-1β antibody (diluted 1 : 100 in PBS; Abcam, UK), and rabbit polyclonal anti-TNFα antibody (diluted 1 : 100 in PBS; Abcam, UK) by sequential incubation. A peroxidase-linked secondary antibody and diaminobenzidine (Sungene Biotech Co., Ltd., Tianjin, China) were used to detect specific immunostaining. The slides were rinsed twice and counterstained with hematoxylin. As negative controls for nonspecific binding of the secondary antibody, sections from the same samples were processed without the primary antibody.

Han L.P., Li C.J., Sun B., Xie Y., Guan Y., Ma Z.J, & Chen L.M. (2016). Protective Effects of Celastrol on Diabetic Liver Injury via TLR4/MyD88/NF-κB Signaling Pathway in Type 2 Diabetic Rats. Journal of Diabetes Research, 2016, 2641248.

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