Revision 3

#6893Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

29

Source/Isotype:

Rabbit IgG

UniProt ID:

#P09429

Entrez-Gene Id:

3146

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

HMGB1 (D3E5) Rabbit mAb recognizes endogenous levels of total HMGB1 protein. This antibody does not cross-react with other HMGB proteins, including HMGB2 and HMGB3.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Chicken, Bovine, Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala137 of human HMGB1 protein.

Background

High mobility group protein B1 (HMGB1) belongs to a family of highly conserved proteins that contain HMG box domains (1,2). All three family members (HMGB1, HMGB2, and HMGB3) contain two HMG box domains and a C-terminal acidic domain. HMGB1 is a widely expressed and highly abundant protein (2). HMGB2 is widely expressed during embryonic development, but is restricted to lymphoid organs and testis in adult animals (3). HMGB3 is only expressed during embryogenesis (4). While expression varies, the biochemical properties of the different family members may be indistinguishable. The HMG box domains facilitate the binding of HMGB proteins to the minor groove of DNA, which results in local bending of the DNA double helix (1,2). HMGB proteins are recruited by and help facilitate the assembly of site-specific DNA binding proteins to their cognate binding sites in chromatin. For example, HMGB1 facilitates the binding of Hox proteins, Oct-1, p53, Rel proteins, and steroid hormone receptor proteins to their target gene promoters (1,2). In addition to their functions in the nucleus, HMGB proteins play a significant role in extracellular signaling associated with inflammation (5,6). HMGB1 is massively released into the extracellular environment during cell necrosis, but not apoptosis. Extracellular HMGB1 "alarms" the innate immune system by acting as a chemoattractant for inflammatory leukocytes, smooth muscle cells, and stem cells, functioning as an immune adjuvant for soluble and particulate antigens, and triggering activation of T cells and dendritic cells. In addition, activated monocytes, macrophages and, dendritic cells also secrete HMGB1, forming a positive feedback loop that results in the release of additional cytokines and neutrophils. Hypoxia has also been shown to cause the release of HMGB1 in the liver, and some studies suggest a role for extracellular HMGB1 in tumor homeostasis (5,6).

  1. Thomas, J.O. and Travers, A.A. (2001) Trends Biochem Sci 26, 167-74.
  2. Müller, S. et al. (2004) J Intern Med 255, 332-43.
  3. Ronfani, L. et al. (2001) Development 128, 1265-73.
  4. Vaccari, T. et al. (1998) Genomics 49, 247-52.
  5. Campana, L. et al. (2008) Curr Opin Immunol 20, 518-23.
  6. Klune, J.R. et al. (2008) Mol Med 14, 476-84.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 3
#6893

HMGB1 (D3E5) Rabbit mAb

Western Blotting Image 1: HMGB1 (D3E5) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using HMGB1 (D3E5) Rabbit mAb.
Western Blotting Image 2: HMGB1 (D3E5) Rabbit mAb Expand Image
Western blot analysis of extracts from the media of mouse bone marrow derived macrophages (mBMDM), untreated (-) or treated with Lipopolysaccharides (LPS) #14011 (50 ng/ml, 4 hr; +) followed by Nigericin (15 μM, 45 min; +), using HMGB1 (D3E5) Rabbit mAb.
Immunohistochemistry Image 1: HMGB1 (D3E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse lung using HMGB1 (D3E5) Rabbit mAb.
Immunohistochemistry Image 2: HMGB1 (D3E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcioma using HMGB1 (D3E5) Rabbit mAb.
Immunohistochemistry Image 3: HMGB1 (D3E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcioma using HMGB1 (D3E5) Rabbit mAb.
Immunohistochemistry Image 4: HMGB1 (D3E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate carcioma using HMGB1 (D3E5) Rabbit mAb.