Anti-Histone H2B (acetyl K20) antibody [EPR859] - ChIP Grade

• Supplier: Abcam
• Catalog: ab177430
• Prices: $99.00
• Sizes: 10 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EPR859
• Applications: IHC-P WB ICC/IF ICC/IF FC ChIP
• Species Reactivities: Mouse, Rat, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Histone H2B aa 1-100 (acetyl K20)
• Description: Rabbit Monoclonal
• Gene: HIST2H2BF
• Conjugate: Unconjugated

Product images

All lanes : Anti-Histone H2B (acetyl K20) antibody [EPR859] - ChIP Grade (ab177430) at 1/10000 dilutionLane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysatesLane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) untreated whole cell lysatesLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1000

All lanes : Anti-Histone H2B (acetyl K20) antibody [EPR859] - ChIP Grade (ab177430) at 1/1000 dilutionLane 1 : NIH/3T3 (Mouse embyro fibroblast cells) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates 10ugLane 2 : NIH/3T3 (Mouse embyro fibroblast cells) untreated whole cell lysates 10ugSecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2B (acetyl K20) with ab177430 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Human colon is observed. Counter stained with Hematoxylin.Negative control: Using PBS instead of primary antibody.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Histone H2B (acetyl K20) with ab177430 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on mouse kidney is observed. Counter stained with Hematoxylin.Negative control: Using PBS instead of primary antibody.

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H2B (acetyl K20) with ab177430 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Rat colon is observed. Counter stained with Hematoxylin.Negative control: Using PBS instead of primary antibody.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells, treated with Trichostatin A (500 ng/ml) for 4 hours or untreated, labeling Histone H2B (acetyl K20)  with ab177430 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line is observed. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows;1. ab177430 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goa

Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells treated with 500ng/ml Trichostatin A for 4 hours, labeling Histone H2B (acetyl K20) with ab177430 at 1/300 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

Chromatin was prepared from HeLa (Human epithelial cells from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab177430 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.