HMGB1 Antibody

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ProSci
Product Code: PSI-7715
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-7715-0.02mg0.02mg£150.00
Quantity:
PSI-7715-0.1mg0.1mg£449.00
Quantity:
Prices exclude any Taxes / VAT

Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Western Blot (WB)

Images

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<strong>Figure 1 KO Validation in HeLa Cells</strong><br> Loading: 10 μg of HeLa WT cell lysates or HMGB1 KO cell lysates. Antibodies: HMGB1 (0.5 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 2 WB Validation in Human, Mouse and Rat Cell Lines </strong><br> Loading: 15 μg of lysate Antibodies: HMGB1 7715, 1 μg/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong> Figure 3 WB Validation in Mouse Tissues</strong><br> Loading: 15 μg of lysate Antibodies: HMGB1 7715, 1 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 4 WB Validation in Rat Tissues</strong><br> Loading: 15 μg of lysate Antibodies: HMGB1 7715, 1 μg/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 5 Immunofluorescence Validation of HMGB1 in HL60 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HL60 cells labeling HMGB1 with 7715 at 20 μg /mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).
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<strong>Figure 6 Immunohistochemistry Validation of HMGB1 in Human Stomach Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded human stomach tissue using anti-HMGB1 antibody (7715) at 1 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 7 Immunohistochemistry Validation of HMGB1 in Mouse Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-HMGB1 antibody (7715) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 8 Immunohistochemistry Validation of HMGB1 in Rat Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-HMGB1 antibody (7715) at 1 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 1 KO Validation in HeLa Cells</strong><br> Loading: 10 μg of HeLa WT cell lysates or HMGB1 KO cell lysates. Antibodies: HMGB1 (0.5 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 WB Validation in Human, Mouse and Rat Cell Lines </strong><br> Loading: 15 μg of lysate Antibodies: HMGB1 7715, 1 μg/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong> Figure 3 WB Validation in Mouse Tissues</strong><br> Loading: 15 μg of lysate Antibodies: HMGB1 7715, 1 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 WB Validation in Rat Tissues</strong><br> Loading: 15 μg of lysate Antibodies: HMGB1 7715, 1 μg/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Immunofluorescence Validation of HMGB1 in HL60 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HL60 cells labeling HMGB1 with 7715 at 20 μg /mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).
<strong>Figure 6 Immunohistochemistry Validation of HMGB1 in Human Stomach Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded human stomach tissue using anti-HMGB1 antibody (7715) at 1 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 7 Immunohistochemistry Validation of HMGB1 in Mouse Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-HMGB1 antibody (7715) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 8 Immunohistochemistry Validation of HMGB1 in Rat Spleen Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-HMGB1 antibody (7715) at 1 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Documents

Further Information

Additional Names:
HMGB1 Antibody: HMG1, HMG3, SBP-1, HMG1, High mobility group protein B1, High mobility group protein 1, HMG-1
Application Note:
WB: 1 μg/mL; IHC: 1-2 μg/mL; IF: 20 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human, mouse, and rat samples; Immunofluorescence in human samples. All other applications and species not yet tested.
Background:
The high-mobility group box 1 (HMGB1) protein was originally identified as a highly conserved nuclear DNA-binding protein that participates in DNA replication, repair and transcriptional regulation of gene expression (1,2). However, HMGB1 has more recently emerged as an extracellularly released mediator of inflammation and repair responses in lipopolysaccharide (LPS)-induced endotoxemia and sepsis (3). HMGB1 has also been suggested to induce secretion of pro-inflammatory cytokines and chemokines by inducing NF-kB signaling downstream of Toll-like receptor (TLR) 2 and TLR4 activation (4).
Background References:
  • Smerdon MJ and Isenberg I. Interactions between the subfractions of calf thymus H1 and nonhistone chromosomal proteins HMG1 and HMG2. Biochemistry 1976; 15:4242-7.
  • Stros. Biocim. Biophys. Acta. 2010; 1799:101-113.
  • Wang et al. Science 1999; 285:248-51.
  • Van Zoelenet al. Shock 2009; 31:280-4.
  • Wei etal. Cell 2021; 184:76-91.
Buffer:
HMGB1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Chicken: (100%), Pig: (100%)
Immunogen:
Anti-HMGB1 antibody (7715) was raised against a peptide corresponding to 19 amino acids near the center of human HMGB1.

The immunogen is located within 70-120 amino acids of HMGB1.
ISOFORMS:
Human HMGB1 has only 1 isoform (215aa, 25kD). Mouse HMGB1 has only 1 isoform (215aa, 25kD). Rat HMGB1 has only 1 isoform (215aa, 25kD). This antibody detects HMGB1 of human, mouse and rat.
NCBI Gene ID #:
3146
NCBI Official Name:
high mobility group box 1
NCBI Official Symbol:
HMGB1
NCBI Organism:
homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 25 kDa

Observed: 25 kDa
Protein Accession #:
NP_002119
Protein GI Number:
4504425
Purification:
HMGB1 antibody is affinity chromatography purified via peptide column.
Research Area:
Innate Immunity
SPECIFICITY:
HMGB1 antibody is human, mouse and rat reactive.
Swissprot #:
P09429
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

KO Validation (Figure 1): Anti-HMGB1 antibodies (7715) specificity was further verified by HMGB1 specific knockout cell lysate. HMGB1 signal was detected in wild type cells, but not in the HMGB1 knockout cells.

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