Anti-HMGB1 antibody [EPR3507]

• Supplier: Abcam
• Catalog: ab79823
• Prices: $404.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EPR3507
• Applications: ICC/IF ICC/IF WB FC IHC-P
• Species Reactivities: Mouse, Rat, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human HMGB1 aa 150 to the C-terminus
• Description: Rabbit Monoclonal
• Gene: HMGB1
• Conjugate: Unconjugated

Product images

All lanes : Anti-HMGB1 antibody [EPR3507] (ab79823) at 1/10000 dilution (unpurified)Lane 1 : SK-BR-3 cell lysateLane 2 : HeLa cell lysateLysates/proteins at 10 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

All lanes : Anti-HMGB1 antibody [EPR3507] (ab79823) at 1/10000 dilution (purified)Lane 1 : SK-BR-3 cell lysateLane 2 : HeLa cell lysateLysates/proteins at 10 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Anti-HMGB1 antibody [EPR3507] (ab79823) at 1/10000 dilution (unpurified) + Rat brain tissue lysate at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Anti-HMGB1 antibody [EPR3507] (ab79823) at 1/10000 dilution (purified) + Rat brain tissue lysate at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

All lanes : Anti-HMGB1 antibody [EPR3507] (ab79823) at 1/50000 dilution (unpurified)Lane 1 : SK-BR-3 cell lysateLane 2 : HeLa cell lysateLane 3 : HepG2 cell lysateLysates/proteins at 10 µg per lane.SecondaryGoat anti-rabbit HRP conjugate at 1/2000 dilution

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling HMGB1 with unpurified ab79823 at 1/350. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling HMGB1 with purified ab79823 at 1/400. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling HMGB1 with unpurified ab79823 at 1/250 dilution.

Immunocytochemsitry/Immunofluorescence analysis of HeLa cells labelling HMGB1 (red) with unpurified ab79823 at 1/350. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

Immunocytochemsitry/Immunofluorescence analysis of HeLa cells labelling HMGB1 (red) with purified ab79823 at 1/400. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

ICC/IF image of unpurified ab79823 stained DU145 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab79823 at 1/1000 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Immunofluorescence analysis of murine RAW 264.7 macrophages, either untreated (upper panel) or treated with LPS (bottom panel). HMGB1 was stained using unpurified ab79823.Cells were fixed in paraformaldehyde, blocked in BSA for 1h, followed by permeabilization in 10% Triton X-100 for 30 min. Samples were incubated with primary antibody overnight at 4°C. An Alexa Fluor® 488-conjugated anti-rabbit IgG was used as the secondary antibody.

Overlay histogram showing HeLa cells stained with unpurified ab79823 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab79823, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.

Flow cytometry analysis of HeLa cells labelling HMGB1 with unpurified ab79823 at 1/30 (red). Cells were fixed with methanol (5 min). A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

Flow cytometry analysis of HeLa cells labelling HMGB1 with purified ab79823 at 1/30 (red). Cells were fixed with methanol (5 min). A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

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