HMGB1/HMG-1 Antibody (951420) [Unconjugated]

Western blot shows lysates of Jurkat human acute T cell leukemia cell line, Hepa 1-6 mouse hepatoma cell line, H4-II-E-C3 rat hepatoma cell line, HEK293 human embryonic kidney cell line, and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Human HMGB1/HMG-1 Monoclonal Antibody (Catalog # MAB16901) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (HAF005). A specific band was detected for HMGB1/HMG-1 at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

HMGB1/HMG-1 was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Rat Anti-Human/Mouse/Rat HMGB1/HMG-1 Monoclonal Antibody (Catalog # MAB16901) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western shows lysates of Jurkat human acute T cell leukemia cell line and Hepa 1‑6 mouse hepatoma cell line, loaded at 0.2 mg/ml. A specific band was detected for HMGB1/HMG‑1 at approximately 35 kDa (as indicated) using 20 µg/mL of Rat Anti-Human/Mouse/Rat HMGB1/HMG‑1 Monoclonal Antibody (Catalog # MAB16901). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, IHC
• Clone: 951420
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

HMGB1/HMG-1 Antibody (951420) [Unconjugated] Summary

• Immunogen: Human HMGB1/HMG-1 synthetic peptide between amino acids Lys50-Ser100
  Accession # P09429
• Specificity: Detects human, mouse, and rat HMGB1/HMG-1 in Western blot.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Rat
• Gene: HMGB1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-25 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for HMGB1/HMG-1 Antibody (951420) [Unconjugated]

• Amphoterin
• high mobility group box 1
• High mobility group protein 1
• high mobility group protein B1
• high-mobility group (nonhistone chromosomal) protein 1
• high-mobility group box 1
• HMG1
• HMG-1
• HMG1DKFZp686A04236
• HMG3
• HMGB1
• SBP-1
• Sulfoglucuronyl carbohydrate binding protein

Background

Human High-mobility group box 1 protein (HMGB1), previously known as HMG-1 or amphoterin, is a member of the high mobility group box family of non-histone chromosomal proteins (1‑3). Human HMGB1 is expressed as a 30 kDa, 215 amino acid (aa) single chain polypeptide containing three domains: two N-terminal globular, 70 aa positively charged DNA-binding domains (HMG boxes A and B), and a negatively charged 30 aa C-terminal region that contains only Asp and Glu (4, 5). Residues 27‑43 and 178‑184 contain a NLS. Posttranslational modifications of the molecule have been reported, with acetylation occurring on as many as 17 lysine residues (6). HMGB1 is expressed at high levels in almost all cells (2, 4). It was originally discovered as a nuclear protein that could bend DNA. Such bending stabilizes nucleosome formation and regulates the expression of select genes upon recruitment by DNA binding proteins (1, 7, 8). It is now known that HMGB1 can also act extracellularly, both as an inflammatory mediator that promotes monocyte migration and cytokine secretion, and as a mediator of T cell-dendritic cell interaction (1, 4, 7, 9, 10). The cytokine activity of HBMG1 is restricted to the HMG B box, (3) while the A box is associated with the helix-loop-helix domain of transcription factors (11). HMBG1 is released in response to cell death and as a secretion product. Although HMBG-1 does not possess a classic signal sequence, it appears to be secreted as an acetylated form via secretory endolysosome exocytosis (6, 12). Once secreted, HMGB1 transduces cellular signals through its high affinity receptor, RAGE and, possibly, TLR2 and TLR4 (1, 3, 4). Human HMGB1 is 100% aa identical to canine HMGB1 and 99% aa identical to mouse, rat, bovine and porcine HMGB1, respectively.

1. Lotze, M.T. and K.J. Tracey (2005) Nat. Rev. Immunol. 5:331.
2. Yang, H. et al. (2005) J. Leukoc. Biol. 78:1.
3. Dumitriu, I.E. et al. (2005) Trends Immunol. 26:381.
4. Degryse, B. and M. de Virgilio (2003) FEBS Lett. 553:11.
5. Wen, L. et al. (1989) Nucleic Acids Res. 17:1197.
6. Bonaldi, T. et al. (2003) EMBO J. 22:5551.
7. Muller, S. et al. (2001) EMBO J. 20:4337.
8. Bustin, M. (1999) Mol. Cell. Biol. 19:5237.
9. Wang, H. et al. (1999) Science. 285:248.
10. Dimitriu, I.E. et al. (2005) J. Immunol. 174:7506.
11. Najima, Y. et al. (2005) J. Biol. Chem. 280:27523.
12. Gardella, S. et al. (2002) EMBO Rep. 3:995.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HMGB1/HMG-1 Antibody (MAB16901)(6)

Publications using MAB16901

• Chi, G;Lu, J;He, T;Wang, Y;Zhou, X;Zhang, Y;Qiu, L; High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-?B pathway following spinal cord injury Scientific reports 2024-09-27 [PMID: 39333662] (In vivo assay, Rat)
• RM Salama, RH Ahmed, AA Farid, BA AbdElSatta, RM AbdelBaset, ME Youssef, SA El Wakeel Gastroprotective effect of dapagliflozin in ethanol-induced gastric lesions in rats: Crosstalk between HMGB1/RAGE/PTX3 and TLR4/MyD88/VEGF/PDGF signaling pathways International immunopharmacology, 2023-01-07;115(0):109686. 2023-01-07 [PMID: 36623411] (Western Blot, Rat)
• Tachpon Techarang, Pitchanee Jariyapong, Parnpen Viriyavejakul, Supattra Glaharn, Charit Srisook, Chuchard Punsawad Protective Effect of an Anti-HMGB-1 Neutralizing Antibody on Hemozoin-Induced Alveolar Epithelial Cell in a Model of Malaria Associated ALI/ARDS Iranian Journal of Parasitology 2021-09-06 [PMID: 34630581]
• D Lian, J Liu, R Han, J Jin, L Zhu, Y Zhang, Y Huang, X Wang, S Xian, Y Chen Kakonein restores diabetes-induced endothelial junction dysfunction via promoting autophagy-mediated NLRP3 inflammasome degradation Journal of Cellular and Molecular Medicine, 2021-06-27;0(0):. 2021-06-27 [PMID: 34180143] (Mouse)
• Xiaowen Bi, Baolin Jiang, Jinyi Zhou, Xirui Fan, Xintong Yan, Juanjuan Liang et al. CBP Bromodomain Inhibition Rescues Mice From Lethal Sepsis Through Blocking HMGB1-Mediated Inflammatory Responses Frontiers in Immunology 2021-02-02 [PMID: 33603756]
• D Wang, K Liu, H Wake, K Teshigawar, S Mori, M Nishibori Anti-high mobility group box-1 (HMGB1) antibody inhibits hemorrhage-induced brain injury and improved neurological deficits in rats Sci Rep, 2017-04-10;7(0):46243. 2017-04-10 [PMID: 28393932] (IHC-P, Rat)

Bioinformatics

• Gene Symbol: HMGB1
• Uniprot:
  • P09429()