HMGB1 Rabbit Polyclonal Antibody

• Catalog Number: orb10828
• Category: Antibodies
• Description: HMGB1 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC
• Predicted Reactivity: Bovine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human HMGB1 (75-170/215aa)
• Concentration: 1mg/ml
• Dilution range: Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 25 kDa
• Target: HMGB1
• UniProt ID: P09429
• RRID: AB_10749907
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: High mobility group protein B1; Amphoterin; High m
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue line): MCF7 (blue). Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (orb10828), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (orb10828), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Positive control: HepG2 cells, Concebtration: 5 µg/10^6 cells, Incubation conditions: Avoid light, 30 minutes on the ice.

Rat splenocytes stained with Anti-HMGB1 Polyclonal Antibody, A488 Conjugated at 1:50.

The figure annotation: The blue histogram is unstained cells. The Orange histogram is cells stained with Rabbit IgG/FITC (orb525556), The green histogram is cells stained with Rabbit Anti-HMGB1/FITC Conjugated antibody. Controls, Positive control: HepG 2 cells, Isotype control: Cell lines treated with Rabbit IgG/FITC (orb525556). instead of the primary antibody to confirm that primary antibody binding is specific. 5 µgin 100 µl 1 X PBS containing 0.5% BSA.