Anti-Zebrafish HMGB1a/b Antibody

Western blot analysis of HMGB1a/b using anti-HMGB1a/b antibody (AZQ6NX86).
Electrophoresis was performed on a 10% SDS-PAGE SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: zebrafish head tissue lysates,
Lane 2: whole female zebrafish tissue lysates,
Lane 3: whole male zebrafish tissue lysates,
Lane 4: zebrafish embryo tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HMGB1a/b antigen affinity purified polyclonal antibody (AZQ6NX86) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for HMGB1a/b at approximately 23 kDa. The expected band size for HMGB1a/b is at 23 kDa.

IHC analysis of HMGB1a/b using anti-HMGB1a/b antibody (AZQ6NX86).
HMGB1a/b was detected in a paraffin-embedded section of zebrafish brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HMGB1a/b Antibody (AZQ6NX86) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of HMGB1a/b using anti-HMGB1a/b antibody (AZQ6NX86).
HMGB1a/b was detected in a paraffin-embedded section of zebrafish brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HMGB1a/b Antibody (AZQ6NX86) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of HMGB1a/b using anti-HMGB1a/b antibody (AZQ6NX86).
HMGB1a/b was detected in a paraffin-embedded section of zebrafish brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HMGB1a/b Antibody (AZQ6NX86) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of HMGB1a/b using anti-HMGB1a/b antibody (AZQ6NX86).
HMGB1a/b was detected in a paraffin-embedded section of zebrafish colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HMGB1a/b Antibody (AZQ6NX86) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of HMGB1a/b using anti-HMGB1a/b antibody (AZQ6NX86).
HMGB1a/b was detected in a paraffin-embedded section of zebrafish eye tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HMGB1a/b Antibody (AZQ6NX86) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.