Anti-Histone H2B Rabbit Monoclonal Antibody

Western blot analysis of Histone H2B/H2BC12 using anti-Histone H2B/H2BC12 antibody (M11068).
Electrophoresis was performed on a 12% 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: human A431 whole cell lysates,
Lane 2: human SIHA whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human U2OS whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat C6 whole cell lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse Nerro-2a whole cell 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-Histone H2B/H2BC12 antigen affinity purified monoclonal antibody (M11068) at a dilution of 1:5000 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 Histone H2B/H2BC12 at approximately 17 kDa. The expected band size for Histone H2B/H2BC12 is at 14 kDa.

IHC analysis of Histone H2B/H2BC12 using anti-Histone H2B/H2BC12 antibody (M11068).
Histone H2B/H2BC12 was detected in a paraffin-embedded section of human breast cancer 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 a dilution of 1:50 rabbit anti-Histone H2B/H2BC12 Antibody (M11068) 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 Histone H2B/H2BC12 using anti-Histone H2B/H2BC12 antibody (M11068).
Histone H2B/H2BC12 was detected in a paraffin-embedded section of human 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 a dilution of 1:50 rabbit anti-Histone H2B/H2BC12 Antibody (M11068) 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 H2B/H2BC12 using anti-H2B/H2BC12 antibody (M11068).
H2B/H2BC12 was detected in a paraffin-embedded section of rat uterus 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 1:50 rabbit anti-H2B/H2BC12 Antibody (M11068) 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 Histone H2B/H2BC12 using anti-Histone H2B/H2BC12 antibody (M11068).
Histone H2B/H2BC12 was detected in a paraffin-embedded section of human colon cnacer 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 a dilution of 1:50 rabbit anti-Histone H2B/H2BC12 Antibody (M11068) 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.

Immunoprecipitating (IP) Histone H2B in K562 whole cell lysate.
Western blot analysis of Histone H2B using anti-Histone H2B antibody (M11068);
Lane 1: K562 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-Histone H2B antibody in K562 whole cell lysate;
Lane 3: anti-Histone H2B antibody (2μg) + K562 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-Histone H2B antigen affinity purified monoclonal antibody (M11068) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for Histone H2B at approximately 17 kDa. The expected band size for Histone H2B is at 14 kDa.

IHC analysis of Histone H2B/H2BC12 using anti-Histone H2B/H2BC12 antibody (M11068).
Histone H2B/H2BC12 was detected in a paraffin-embedded section of mouse ovary 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 a dilution of 1:50 rabbit anti-Histone H2B/H2BC12 Antibody (M11068) 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.