| Field | Specification |
|---|---|
| Target | |
| Alternative names | High mobility group protein B1; High mobility group protein 1; HMG-1; HMGB1; HMG1 |
| UniProt # | |
| Host | |
| Clone | |
| Clonality | |
| Isotype | |
| Reactivity | |
| Applications | |
| Immunogen | A synthetic peptide corresponding to a sequence at the C-terminus of human HMGB1, identical to the related mouse and rat sequences. |
| Molecular weight | |
| Reconstitution | |
| Cellular localization | |
| Concentration | |
| Form | Lyophilized |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
Clone 5H3 is a mouse monoclonal antibody that recognizes high mobility group protein B1 (gene HMGB1) in human, monkey, mouse and rat samples and is validated for Western blot, IHC and flow cytometry. The predicted molecular weight is 24.9 kDa, and the supplier reports an observed band at about 25 kDa.
HMGB1 is an abundant non-histone chromatin protein that bends DNA and assists transcription, DNA repair and V(D)J recombination. When released from dying or activated cells it acts as a danger signal (DAMP) that engages receptors such as RAGE and TLR4, and it is widely measured in studies of inflammation, sepsis, tissue injury and cell death.
| Target | High mobility group protein B1 (gene HMGB1; UniProt P09429, human) |
|---|---|
| Clone | 5H3 |
| Host / Clonality / Isotype | Mouse / Monoclonal / Mouse IgG2b |
| Reactivity | Human, Monkey, Mouse, Rat |
| Form | Lyophilized |
| Formulation | Per vial: 4 mg trehalose; 0.9 mg NaCl; 0.2 mg Na2HPO4; 0.05 mg NaN3 |
| Calculated MW | 24.9 kDa |
| Observed MW | 25 kDa |
| Storage | As supplied: −20 °C for up to 12 months from receipt. After reconstitution: 4 °C for up to 1 month, or aliquot and keep at −20 °C for up to 6 months. Avoid repeated freeze–thaw cycles. |
Validated Applications
| Western blot | 0.1–0.5 µg/mL (Human, Mouse, Rat, Monkey) |
|---|---|
| IHC (paraffin sections) | 0.5–1 µg/mL (Human, Mouse, Rat) |
| Flow cytometry (fixed cells) | 1–3 µg/1×106 cells (Human) |
Samples with a confirmed band (WB): human HepG2 cells, human CCRF-CEM cells, monkey COS-7 cells, human SW620 cells, human THP-1 cells, rat PC-12 cells, rat RH35 cells, mouse NIH/3T3 cells, mouse RAW264.7- WT cells, mouse RAW264.7-Hmgb1 KO cells.
Recommended loading (WB): 20–40 µg of total protein per lane.
Conditions in the example images (WB): 5–20% gradient SDS-PAGE under reducing conditions, 50 µg lysate per lane, transfer to nitrocellulose membrane, blocking in 5% non-fat milk, primary antibody at 0.5 µg/mL overnight at 4 °C, HRP-conjugated secondary antibody at 1:10000, ECL detection.
Samples with confirmed staining (IHC): rat brain tissue, human intestinal cancer tissue, human mammary cancer tissue, human lung cancer tissue, mouse brain tissue.
Recommended antigen retrieval: heat-mediated, in TE buffer (pH 9.0); citrate buffer (pH 6.0) can be used instead.
Conditions in the example images (IHC): heat-mediated antigen retrieval in citrate buffer (pH 6).
Samples with a confirmed signal (flow cytometry): SiHa cells.
Immunogen
Synthetic peptide from the C-terminal region of human HMGB1. The peptide sequence is conserved in mouse and rat.
Reactivity Notes
The supplier lists reactivity with human, monkey, mouse and rat. UniProt describes high mobility group protein B1 as secreted and also located in the nucleus, chromosome and cytoplasm. Tissue expression noted by UniProt (human): Ubiquitous. The samples tested by the supplier (listed above) are a practical starting point for positive controls.
Safety
Customization & Add-ons: Can’t find the antibody you need—or require a custom format for your assay? We can help you source the best match or support custom antibody solutions for diverse research needs, including species and isotype selection, conjugations and labeling (e.g., HRP/AP, biotin, fluorophores), purification grade options (Protein A/G, affinity purified), formulation preferences (buffer selection, carrier-free, glycerol-free), custom concentrations and aliquoting, low-endotoxin options for cell-based work, and application-focused QC/validation support (project dependent). Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services for additional support—our team will follow up with feasibility details and next steps.
Liucheng Li et al. (2014) Total extract of Yupingfeng attenuates bleomycin-induced pulmonary fibrosis in rats. Phytomedicine. 10.1016/j.phymed.2014.10.011
