HMGB1 Antibody

• Catalog Number: orb1239440
• Category: Antibodies
• Description: HMGB1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, WB
• Predicted Reactivity: Gallus, Porcine
• Reactivity: Human, Mouse, Rat
• 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.
• Concentration: 1 mg/mL
• Dilution range: 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.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 25 kDa Observed: 25 kDa
• Target: HMGB1
• UniProt ID: P09429
• NCBI: NP_002119
• Storage: Antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures
• Buffer/Preservatives: HMGB1 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: HMGB1 Antibody: HMG1, HMG3, SBP-1, HMG1, High mobi
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

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.