Cultured cells were fixed with 2% PFA (Electron Microscopy Sciences) in PBS (Crystalgen) for 15 min after washing with HBSS and quenched with 0.1 M glycine (Sigma-Aldrich) in PBS. For intracellular staining, the cells were treated with 0.1% Triton X-100 (Fisher Scientific) in 1% BSA (Roche) in PBS for 5 min. Cells were blocked with 10% goat serum (Sigma-Aldrich) in PBS for 1 h, and then stained for 1 h with primary antibody diluted in 3% BSA in PBS. After washing, cells were stained with fluorescent-conjugated secondary antibody for 30 min. Cells were washed with PBS, stained with DAPI, and mounted with fluorescent mounting medium (Dako). All procedures were performed at room temperature. Anti-MAP2 antibody (Sigma-Aldrich), anti-postsynaptic density protein 95 (PSD-95) antibody (Millipore), anti-NMDAR subunit 1 extracellular (GluN1) antibody (Alomone Labs), antiHMGB1 antibody (Novus), and anti-C1q antibody (Abcam) were used for immunofluorescence staining. Secondary antibodies, Alexa Fluor Goat anti-Mouse antibody and Goat anti-Rabbit antibody (Life Technologies) were used following primary antibody staining.
Anti hmgb1 antibody
Manufactured by Novus Biologicals
Sourced in United States
The Anti-HMGB1 antibody is a laboratory reagent used for the detection and analysis of HMGB1 (High Mobility Group Box 1) protein in various biological samples. HMGB1 is a nuclear protein involved in DNA binding and transcriptional regulation. This antibody can be used in techniques such as Western blotting, immunohistochemistry, and ELISA to study the expression and localization of HMGB1 in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescent mounting medium, by Agilent Technologies (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Anti c1q antibody, by Abcam (1 mentions)
Anti map2 antibody, by Merck Group (1 mentions)
Triton x 100, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Roche (1 mentions)
Anti rabbit horseradish peroxidase linked whole antibody, by GE Healthcare (1 mentions)
Anti 4 hne antibody, by Abcam (1 mentions)
Anti actin antibody, by Merck Group (1 mentions)
Enhanced chemi luminescence (ecl), by Merck Group (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
3 protocols using anti hmgb1 antibody
1
Immunofluorescence Staining of Cultured Cells
2
Modulating HMGB1 for Nerve Regeneration
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To investigate the associations among HMGB1, the inflammatory reaction, and nerve regeneration after injury, the anti-HMGB1 antibody (Novus Biologicals, USA) was injected intraperitoneally just after the NTx. To determine if there is a dose-dependent effect of the drug, low (25 μg in 0.5 ml saline) and high (50 μg in 0.5 ml saline) doses of the anti-HMGB1 antibody were used, making them comparable to a dose used in a previous study [25 (link)]. We also injected IgG (50 μg in 0.5 ml saline) intraperitoneally in a group of control animals. We collected data from six mice for each of the three treatment groups and each of the four recovery time points (days 5, 14, 42, and 100) for a total of 72 mice (6 mice × 3 doses × 4 recovery points).
3
Immunoblotting Analysis of Cochlear Proteins
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Aliquots of cochlea homogenates were separated on 8% sodium dodecyl sulfate (SDS) polyacrylamide gels, transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA, USA), blocked with 5% skimmed milk in TBST (0.2 M Tris-base, 1.37 M NaCl, and 0.1% Tween 20), probed with the indicated primary antibody at 4 °C overnight, washed with TBST, and incubated with anti-rabbit horseradish peroxidase-linked whole antibody (1:10,000; GE Healthcare, Chicago, IL, USA) for 1 h at RT. The immunoreactive bands were stained using a light emitting nonradioactive method (ECL; Millipore). The specific primary antibodies were anti-HMGB1 antibody (1:1000; Novus Biologicals, Littleton, CO, USA), anti-4-HNE antibody (1:1000; Abcam, Cambridge, UK), and anti-actin antibody (1:1000; Millipore, Burlington, MA, USA).
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