Loading...

Purified anti-HMGB1 Antibody

Pricing & Availability
Clone
A15164F (See other available formats)
Regulatory Status
RUO
Other Names
High mobility group protein B1, High mobility group protein 1, HMG-1,
Isotype
Mouse IgG2a, κ
Ave. Rating
Submit a Review
Product Citations
publications
A. A15164F_PURE_WB_1_050125
Whole cell extracts (15 µg total protein) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-HMGB1 (clone A15164F) overnight at 4 °C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 643807) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • A. A15164F_PURE_WB_1_050125
    Whole cell extracts (15 µg total protein) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-HMGB1 (clone A15164F) overnight at 4 °C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 643807) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • b. A15164F_PURE_WB_2_050125
    Whole cell extracts (15 µg total protein) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-HMGB1 (clone A15164F) overnight at 4 °C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-mouse IgG (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 607904) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • c. A15164F_PURE_ICC_3_050125
    HeLa cells were fixed with Fixation Buffer (Cat. No. 420801), then permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained without (panel A and C) and with (panel B and D) Purified anti-HMGB1 (clone A15164F), followed by incubation with Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322) (red) for 1 hour at room temperature. Cells were co-stained with Alexa Fluor® 488 anti-Tubulin-α (Cat. No. 627906) (green) and nuclei were counterstained with DAPI (Cat. No. 422801) (blue). The images were captured on a Revvity Operetta CLS™ High Content Analysis System with a 63X objective and merged (panel C and D). Scale bar: 50 µm
  • d. A15164F_PURE_ICFC_4_070225
    MCF7 cells (filled histogram, high-expressing cell line) and HeLa cells (open histogram, low-expressing cell line) were fixed with Fixation Buffer (Cat. No. 420801), permeabilized using True-Phos™ Perm Buffer (Cat. No. 425401) and intracellularly stained with Purified anti-HMGB1 (clone A15164F), or Purified Mouse IgG2a, κ Isotype Control (open histogram, dashed line, representative of both positive and negative cell lines) (Cat. No. 400202) followed by Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322).
Cat # Size Price Quantity Save
641251 25 µg ¥36,300
641252 100 µg ¥97,900
Description

High mobility group protein B1 (HMGB1) belongs to a family of highly conserved proteins that contain HMG box domains. Human HMGB1 is 100% identical to the sequence of canine HMGB1, and 99% 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 but is now known to also be secreted and play a significant role in extracellular signaling as a proinflammatory cytokine. It acts as an inflammatory mediator that promotes monocyte migration, cytokine secretion, and mediate 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

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Full length recombinant mouse HMGB1
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 10.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application. For flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Additional Product Notes

For use of this antibody in immunocytochemistry (ICC), it is recommended to fix/permeabilize with either of the following:-Fixation buffer (Cat. No. 420801) with 0.5 % Triton X-100-Fixation buffer (Cat. No. 420801) with 100% ice-cold methanol.

For use of this antibody in intracellular flow cytometry (ICFC), it is recommended to fix/permeabilize with either of the following:

-True-Phos™ Perm Buffer (Cat. No. 425401).
-True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401).

*Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) is not recommended with this antibody.

RRID
AB_3716968 (BioLegend Cat. No. 641251)
AB_3716968 (BioLegend Cat. No. 641252)

Antigen Details

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

Nucleus

Interaction
CXCL12, RAG Complex, BECN1, KPNA1
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

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 05/01/2025

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.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
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.

ProductsHere
Login / Register
Remember me
Forgot your password? Reset password?
Create an Account