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Description
Research Area
Images & Validation
−| Tested Applications | ELISA, IF, IHC-P, WB |
|---|---|
| Reactivity | Human, Mouse, Rat |
| Predicted Reactivity | Gallus, Porcine |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Clonality | Polyclonal |
| Isotype | IgG |
| Immunogen | Anti-HMGB1 antibody (orb1239440) 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. |
| Target | HMGB1 |
| Molecular Weight | Predicted: 25 kDa Observed: 25 kDa |
| Purification | HMGB1 antibody is affinity chromatography purified via peptide column. |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures |
|---|---|
| Form/Appearance | Liquid |
| Buffer/Preservatives | HMGB1 Antibody is supplied in PBS containing 0.02% sodium azide. |
| Concentration | 1 mg/mL |
| Expiration Date | 12 months from date of receipt. |
| Disclaimer | For research use only |
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KO Validation in HeLa Cells. Loading: 10 µg of HeLa WT cell lysates or HMGB1 KO cell lysates. Antibodies: HMGB1 (0.5 µg/mL) and beta-actin orb1240312 (1 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

WB Validation in Human, Mouse and Rat Cell Lines. Loading: 15 µg of lysate Antibodies: HMGB1 orb1239440, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

WB Validation in Mouse Tissues. Loading: 15 µg of lysate Antibodies: HMGB1 orb1239440, 1 µ g/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

WB Validation in Rat Tissues. Loading: 15 µg of lysate Antibodies: HMGB1 orb1239440, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Immunofluorescence Validation of HMGB1 in HL60 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed HL60 cells labeling HMGB1 with orb1239440 at 20 µg /mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunohistochemistry Validation of HMGB1 in Human Stomach Tissue. Immunohistochemical analysis of paraffin-embedded human stomach tissue using anti-HMGB1 antibody (orb1239440) 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.

Immunohistochemistry Validation of HMGB1 in Mouse Spleen Tissue. Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-HMGB1 antibody (orb1239440) 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.

Immunohistochemistry Validation of HMGB1 in Rat Spleen Tissue. Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-HMGB1 antibody (orb1239440) 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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HMGB1 Antibody (orb1239440)
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