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Anti-HMGB1 Antibody EPR3507
Also for HMGB1 (NM_002128)
|A synthetic peptide corresponding to residues in human HMGB1 was used as an immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||ICC/IF: Use at an assay dependent concentration; WB: 1:10000 - 1:50000; IP: 1:20; FC: 1:20; IHC-P: 1:250 - 1:500
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Homo sapiens high mobility group box 1 (HMGB1)|
|HMG1; HMG3; SBP-1|
|High-mobility group box 1 protein (HMGB1) is a nuclear DNA-binding protein, which also functions as a pleiotropic cytokine, implicated in the pathology of several different immune-mediated diseases (1). As a nuclear protein, HMGB1 stabilizes nucleosomes and allows bending of DNA that facilitates gene transcription. Recent studies identified extracellular HMGB1 as a potent macrophage-activating factor, signaling via the receptor for advanced glycation end-products to induce inflammatory responses. HMGB1 mediates endotoxin lethality, acute lung injury, arthritis induction, activation of macrophages, smooth muscle cell chemotaxis, and epithelial cell barrier dysfunction (2). |
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Western blot - HMGB1 antibody [EPR3507]; All lanes : Anti-HMGB1 antibody [EPR3507] at 1/50000 dilution.Lane 1 : SKBR-3 cell lysate.Lane 2 : HeLa cell lysate.Lane 3 : HepG2 cell lysate.Lysates/proteins at 10 µg per lane.Secondary.Goat anti-rabbit HRP conjugate at 1/2000 dilution.Predicted band size : 25 kDa.Observed band size : 25 kDa.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HMGB1 antibody [EPR3507]; ab79823, at 1/250 dilution, staining HMGB1 in paraffin-embedded human kidney tissue by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Anti-HMGB1 antibody [EPR3507]; Immunofluorescence analysis of murine RAW 264.7 macrophages, either untreated (upper panel) or treated with LPS (bottom panel). HMGB1 was stained using ab79823. Cells were fixed in paraformaldehyde, blocked in BSA for 1h, followed by permeabilization in 10% Triton X-100 for 30 min. Samples were incubated with primary antibody overnight at 4°C. An AlexaFluor?488-conjugated anti-rabbit IgG was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-HMGB1 antibody [EPR3507]; ICC/IF image of ab79823 stained DU145 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab79823 at 1/1000 dilution overnight at +4°C. The secondary antibody (green)was DyLight? 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43uM.
Flow Cytometry - Anti-HMGB1 antibody [EPR3507]; Overlay histogram showing HeLa cells stained with ab79823 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.