Anti-High-Mobility Group Protein 1 (HMGB1) Antibody (1F3)
Our Anti-High-mobility group protein 1 (HMGP-1) mouse monoclonal primary antibody detects bovine, human, mouse, other mammals (predicted), pig, and rat High-mobility group protein 1 (HMGP-1), and is IgG. It is validated for use in FC, ICC, WB.
A: HeLa cells stained with M-1702-100 (red), chicken polyclonal antibody to Vimentin (C-1409-50, green) and DNA (blue). The M-1702-100 antibody reveals strong nuclear staining which overlaps with the DNA stain. B: Blot of crude HeLa cell extract stained with M-1702-100. HMGB1 runs at an apparent molecular weight of 25 kDa. C: Analysis of HMGB1 expression in human euroblastoma SH-SY5Y cell line by Flow Cytometry. Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 1% BSA, Primary antibody: Mouse Monoclonal antibody to HMGB1 (cat # M-1702-100, 2μg per ~10^6 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 fold dilution) with incubation for 20 minutes in dark at room temperature. Non-specific Control IgG, clone X63 (cat # M-1249-200) was used as negative control under same conditions (black dashed). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols.
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SKU: M-1702-100
Product Details
High-mobility group protein 1 (HMGP-1)
High-mobility group proteins were named originally since they are abundand relatively low molecular weight proteins which run quickly on SDS-PAGE gels. High-mobility group protein box 1 (HMGB1, Amphoterin) is one of these. The "bx" in the name refers to the so-called high mobility group (HMG) box, a compact domain involved in DNA binding and protein-protein interactions. the HMGB1 molecule has two of these HMG domains. The protein is alslo called amphoterin, this name being derived from the presence of two highly charged regions in the molecule, a relatively neutrally charged N-terminus and a very negatively charged C-terminus. In fact the molecule is very unusually charged throughout, the human sequence consisting of 16.7% Glutamic acid, 9.3% Aspartic acid, 20% lysine and 9.3% Arginine. HMGB1 can bind Toll like receptor 4 (TLR4) and the Receptor for Advanced Glycation End products (RAGE). TLRs are components of the innate immune system, first recognized as a family of receptors which recognize "Pathogen Associated Molecular Pattern molecules (PAMPs). PAMPs are common components of bacteria and when TLRs bind these a strong inflammatory response is activated. More recently it has been recognized that TLRs can also be activated by Damage Associated Molecular Pattern molecules (DAMPs), which are endogenous substances released from damaged and diseased cells which also bind to TLR family receptors and also activate inflammation. HMGB1 is such a DAMP, binding to TLR4, and much evidence suggests that HMGB1 is a strong activator of inflammation. Interestingly, HMGB1 is released by necrotic cells but not by apoptotic cells (1).
IgG
Monoclonal
1F3
IgG2b
Flow, ICC, WB
Mouse
25 kDa
Human full length recombinant human HMGB1 protein expressed in and purified from E. coli.
Human
Bovine, Human, Mouse, Pig, Rat
Spin vial briefly before opening. Reconstitute with 100 µL sterile-filtered, ultrapure water to achieve a 1 mg/mL concentration. Centrifuge to remove any insoluble material. After reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles.
Lyophilized
Protein G purified
Lyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and sodium azide
WB: 1:1000-1:2000
ICC: 1:500-1:1000
ICC: 1:500-1:1000
Use 2 ug/10^6 cells for Flow cytometry.
Unconjugated
The antibody reacts with a band at ~25 kDa by Western blot on HeLa cell extract. It has also been used successfully for immunocytochemistry showing strong nuclear staining.
For research use only.
United States
12 months after date of receipt (unopened vial).
25°C (ambient)

