Western blot analysis was used to detect protein expression in the ischemic tissue, as described by us in previous work21 (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.
Rabbit polyclonal anti il 1β antibody
Manufactured by Abcam
Sourced in United Kingdom
Rabbit polyclonal anti-IL-1β antibody is a laboratory reagent used for the detection and quantification of interleukin-1 beta (IL-1β) in various biological samples. It is a polyclonal antibody raised in rabbits against the IL-1β protein.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit polyclonal anti tnfα antibody, by Abcam (2 mentions)
Goat anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Polyclonal rabbit anti bcl 2, by Abcam (1 mentions)
Rabbit polyclonal anti bax antibody, by Abcam (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
Protein extraction kit, by Beyotime (1 mentions)
Laemmli sample buffer, by Bio-Rad (1 mentions)
Cresyl violet, by Merck Group (1 mentions)
Hoechst 33258, by Beyotime (1 mentions)
Naltrindole, by Bio-Techne (1 mentions)
Mouse monoclonal anti β actin antibody, by Merck Group (1 mentions)
Tan67, by Bio-Techne (1 mentions)
Rabbit anti iba 1 ab, by Fujifilm (1 mentions)
Mouse anti hmgb1 mab, by R&D Systems (1 mentions)
Mouse monoclonal anti cd68 antibody, by Abcam (1 mentions)
Rabbit polyclonal anti nf κb p65 antibody, by Abcam (1 mentions)
Mouse monoclonal anti tlr4 antibody, by Abcam (1 mentions)
4 protocols using rabbit polyclonal anti il 1β antibody
1
Quantifying Protein Expression in Ischemic Tissue
2
Neuroinflammation and Opioid Receptor Signaling
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Tan67 and Naltrindole (NTI) were purchased from Tocris bioscience (Cat: 0921, 0740). Cresyl violet (CV) was purchased from Sigma-Aldrich (Cat: C5402). Hoechst 33258, BCA assay kit (BSA standard solution) and the protein extraction kit were purchased from Beyotime Biotechnology (Cat: C1017, P0010, and P0013B). Mouse monoclonal anti-IL-10 (A-2) antibody was purchased from Santa Cruz Biotechnology (Cat: SC-365858). Rabbit polyclonal anti-IL-1β antibody was obtained from Abcam Inc (Cat: ab9722). Rabbit polyclonal anti-DOR antibody was obtained from Millipore Sigma (Cat: AB1560). Mouse monoclonal anti-β-actin antibody was obtained from Sigma-Aldrich (Cat: A5441). Alexa Fluor 647-conjugated Affinipure Donkey Anti-Rabbit IgG (H + L), Alexa Fluor 488-conjugated Affinipure Donkey Anti-Mouse IgG (H + L), Peroxidase AffiniPure Goat Anti-Rabbit IgG (H + L), and Peroxidase AffiniPure Goat Anti-Mouse IgG (H + L) are Jackson Immuno Research products (Cat: 711-605-152, 715-545-150, 111-035-003, and 115-035-003). Laemmli sample buffer is from Bio-rad (Cat: 1610747).
3
Immunofluorescence Staining of Brain Sections
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For immunofluorescence staining (Fig. 9 , protocol #1c), the mice were divided into 4 groups with 8 mice in each group. Mice were treated as described in protocol #1 histochemical staining section above. Paraffin-embedded brain sections (5 μm) were stained using a mouse anti-HMGB1 mAb (R&D Systems Inc.), rabbit anti-MAP2 pAb (Santa Cruz Biotechnology Inc.) for HMGB1 detection. For astrocytes and microglia cells staining rabbit anti-GFAP pAb (Abcam, Cambridge, UK), rabbit anti-Iba-1 Ab (Wako, Osaka, Japan) were used. Rabbit polyclonal anti-IL-1β antibody (Abcam, Cambridge, UK) were used for IL-1β detection as the primary antibody. See details in the Supplemental Methods.
4
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.
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