HMGB1/HMG-1 Antibody (1F3) - BSA Free

Western Blot: HMGB1/HMG-1 Antibody (1F3) [NBP2-25148] - Analysis of lysates from different cell lines probed with mouse mAb to high mobility group protein B1, (HMGB1), NBP2-25148, dilution 1:2,000: [1] protein standard, [2] NIH-3T3, [3] C6, [4] HEK293, [5] HeLa, and [6] SH-SY5Y. The 25kDa band revealed by MCA-1F3 antibody, corresponds to HMGB1 protein.

Immunocytochemistry/Immunofluorescence: HMGB1/HMG-1 Antibody (1F3) [NBP2-25148] - Analysis of of HeLa cells stained with mouse mAb to HMGB1,NBP2-25148, dilution 1:2,000 in red and costained with rabbit pAb to GAPDH, dilution 1:2,000 in green. MCA-1F3 antibody stains the chromatin binding protein HMGB1, which is localized in the nuclei.

Flow Cytometry: HMGB1/HMG-1 Antibody (1F3) [NBP2-25148] - Intracellular flow cytometric staining of 1 x 10^6 MCF-7 cells using HMGB1 antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.

Immunocytochemistry/Immunofluorescence: HMGB1/HMG-1 Antibody (1F3) [NBP2-25148] - HeLa cells grown in tissue culture and stained with NBP2-25148 (red), chicken polyclonal antibody to Vimentin NB300-223 (green) and DNA (blue). NBP2-25148 reveals strong nuclear staining which overlaps with the DNA stain.

Simple Western: HMGB1/HMG-1 Antibody (1F3) [NBP2-25148] - Simple Western lane view shows a specific band for HMGB1 in 0.05 mg/ml of Jurkat lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, CyTOF-ready
• Clone: 1F3
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

HMGB1/HMG-1 Antibody (1F3) - BSA Free Summary

• Immunogen: Full length recombinant human HMGB1 protein expressed in and purified form E. coli. [UniProt# P09429]
• Localization: Nucleus. Chromosome.
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: HMGB1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 1 ug per million cells
  • Immunocytochemistry/ Immunofluorescence 1:1000
  • Immunohistochemistry 1:1000
  • Simple Western 1:500
  • Western Blot 1:1000-1:2000
• Application Notes: This HMGB1 (1F3) antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry, and Western Blot, where a band is seen at approximately 25 kDa. This antibody is CyTOF ready.
  
  In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in Jurkat lysate 0.05 mg/mL, separated by Size, antibody dilution of 1:500, apparent MW was 36 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Theoretical MW: 25 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 50% PBS, 50% glycerol
• Preservative: 0.035% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for HMGB1/HMG-1 Antibody (1F3) - BSA Free

• 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

HMGB1 (High-mobility group protein box 1) is part of the high-mobility group proteins which were originally named this way since they are abundant, relatively low molecular weight proteins which run quickly on SDS-PAGE gels. The "box" in the name refers to a compact domain involved in DNA binding and protein-protein interactions. The HMGB1 molecule has two of these HMG domains. The protein is also 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.

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 (NBP2-25148)(2)

Publications using NBP2-25148

• Tonner H, Hunn S, Auler N et al. A Monoclonal Anti-HMGB1 Antibody Attenuates Neurodegeneration in an Experimental Animal Model of Glaucoma International journal of molecular sciences 2022-04-07 [PMID: 35456925] (In Vivo, Rat)
• Nestor J, Arinuma Y, Huerta TS et al. Lupus antibodies induce behavioral changes mediated by microglia and blocked by ACE inhibitors. J. Exp. Med. 2018-09-05 [PMID: 30185634] (ICC/IF, Mouse)

Bioinformatics

• Gene Symbol: HMGB1
• Entrez:
  • 3146(Human)
• Uniprot:
  • P09429()