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<strong>Figure 1 Immunofluorescence Validation of HMGB1 In HeLa Cells</strong><br>Immunofluorescent analysis of PFA-fixed HeLa cells labeling HMGB1 with 7715 at 20 ?g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and Hoechst staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
<strong>Figure 2 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 ?-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 3 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 4 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 5 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 6 Immunohistochemistry Validation of HMGB1 in Human Kidney Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded human kidney 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 7 Immunohistochemistry Validation of HMGB1 in Mouse Kidney Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse kidney 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 Kidney Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded rat kidney 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 9 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.
Figure 1 Immunofluorescence Validation of HMGB1 In HeLa Cells
Immunofluorescent analysis of PFA-fixed HeLa cells labeling HMGB1 with 7715 at 20 ?g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and Hoechst staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
Trial Size

HMGB1 Antibody

CATALOG NUMBER: 7715

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
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.
Specificity:
HMGB1 antibody is human, mouse and rat reactive.
Positive Control 1:
Cat. No. 1402- Mouse Lung Lysate
Positive Control 2:
Cat. No. 10-101 - Human Lung Tissue Slide
Predicted Molecular Weight:
Predicted: 25 kDa
Observed: 25 kDa
Validation:

KO Validation (Figure 2): 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.

.
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.
Purification:
HMGB1 antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
HMGB1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
HMGB1 antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
HMGB1
Additional Names:
HMGB1 Antibody: HMG1, HMG3, SBP-1, HMG1, High mobility group protein B1, High mobility group protein 1, HMG-1
Protein Accession Number:
NP_002119
Protein Gi Number:
4504425
Ncbi Gene Id Number:
3146
User Note:
Optimal dilutions for each application to be determined by the researcher.
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 Reference 1:
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.
Background Reference 2:
Stros. Biocim. Biophys. Acta. 2010; 1799:101-113.
Background Reference 3:
Wang et al. Science 1999; 285:248-51.
Background Reference 4:
Van Zoelenet al. Shock 2009; 31:280-4.
Background Reference 5:
Wei etal. Cell 2021; 184:76-91.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

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.

CATALOG NUMBER:

7715

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
Shipping Info

$65 Standard Overnight flat rate for as many products shipped within the United States.

$35 Dry Ice Shipment fee may be required on some items.

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