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Ultra-LEAF™ Purified anti-HMGB1 Antibody

Pricing & Availability
Clone
3E8 (See other available formats)
Regulatory Status
RUO
Other Names
High mobility group protein 1, High mobility group protein B1, Sulfoglucuronyl carbohydrate binding protein
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
3E8_ULEAF_HMGB1_Antibody_1_022218
Whole cell lysates (15 µg protein) from HeLa and Raw264.7 cells were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:1000 (0.5 µg /mL) Ultra-LEAF™ Purified anti-HMGB1 Antibody, clone 3E8. Proteins were visualized using chemiluminescence detection by incubating with 1:3000 dilution of HRP goat anti-mouse-IgG secondary antibody (Cat. No.405306) . 1:2000 dilution of Direct-Blot™ HRP anti-β-actin Antibody (Cat. No. 643807) was used as a loading control (lower). Lane M: MW ladder.
  • 3E8_ULEAF_HMGB1_Antibody_1_022218
    Whole cell lysates (15 µg protein) from HeLa and Raw264.7 cells were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:1000 (0.5 µg /mL) Ultra-LEAF™ Purified anti-HMGB1 Antibody, clone 3E8. Proteins were visualized using chemiluminescence detection by incubating with 1:3000 dilution of HRP goat anti-mouse-IgG secondary antibody (Cat. No.405306) . 1:2000 dilution of Direct-Blot™ HRP anti-β-actin Antibody (Cat. No. 643807) was used as a loading control (lower). Lane M: MW ladder.
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651413 100 µg £185
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651414 1 mg £490
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Description

High mobility group protein B1 (HMGB1) belongs to a family of highly conserved proteins that contain HMG box domains. Human HMGB1 is expressed as a 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. Human HMGB1 is 100% aa identical to canine HMGB1 and 99% aa identical to mouse, rat, bovine and porcine HMGB1.

HMGB1 is a widely expressed and highly abundant protein. 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. It is now known that HMGB1 also plays a significant role in extracellular signaling associated with inflammation. HMGB1 is massively released into the extracellular environment during cell necrosis.  It acts as an inflammatory mediator that promotes monocyte migration and cytokine secretion, and as a mediator of T cell-dendritic cell interaction. 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.  The cytokine activity of HMGB1 is restricted to the HMG B box, while the A box is associated with the helix-loop-helix domain of transcription factors. Although HMGB1 does not possess a classic signal sequence, it appears to be secreted as an acetylated form via secretory endolysosome exocytosis. Once secreted, HMGB1 transduces cellular signals through its high affinity receptor, RAGE and, possibly, TLR2 and TLR4.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant human HMGB1 with GST tag. A universal T cell epitope from a Mycobacterium tuberculosis antigen was introduced into the C-terminus of HMGB1 to increase the immunogenicity.
Formulation
0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.
Endotoxin Level
Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.
Preparation
The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration
The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.
Application

WB - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.5 - 1.0 µg per ml (1:500 - 1:1000 dilution). It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone 3E8 has been reported to protect mice from LPS induced sepsis1. Additional reported applications (for relevant formats) include: neutralization1. The LEAF™ or Ultra-LEAF™ purified antibody is recommended for functional assays (contact our custom solutions team). 

Application References

(PubMed link indicates BioLegend citation)
  1. Zhou H, et al. 2009. PLos One 4:e6087. (Neut)
Product Citations
  1. de Mingo Pulido , et al. 2021. Immunity. 54(6):1154-1167.e7. PubMed
RRID
AB_2728487 (BioLegend Cat. No. 651413)
AB_2728488 (BioLegend Cat. No. 651414)

Antigen Details

Structure
215 amino acids with predicted molecular weight of 25 kD.
Distribution

Nucleus

Function
DNA binding protein that associates with chromatin and has the ability to bend DNA. Binds single-stranded DNA preferentially. Involved in V(D)J recombination by acting as a cofactor of the RAG complex.
Interaction
Component of the RAG complex composed of core components RAG1 and RAG2.
Cell Type
B cells
Biology Area
Cell Biology, Immunology, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References

1. Thomas JO and Travers AA. 2001. Trends Biochem. Sci. 26:167.
2. Müller S, et al. 2004. J. Intern. Med. 255:332.
3. Campana L, et al. 2008. Curr. Opin. Immunol. 20:518.
4. Klune JR, et al. 2008. Mol. Med. 14:476.
5. Dumitriu IE, et al. 2005. Trends Immunol. 26:381.
6. Bonaldi T, et al. 2003. EMBO J. 22:5551.

Gene ID
3146 View all products for this Gene ID
UniProt
View information about HMGB1 on UniProt.org

Related FAQs

Do you guarantee that your antibodies are totally pathogen free?

BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.

Does BioLegend test each Ultra-LEAF™ antibody by functional assay?

No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.

Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?

No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.

Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?

We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.

Go To Top Version: 1    Revision Date: 02/22/2018

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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