Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade

• Supplier: Abcam
• Catalog: ab176840
• Prices: $99.00
• Sizes: 10 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EPR17899
• Applications: ICC/IF ICC/IF ChIP IHC-P WB DB
• Species Reactivities: Mouse, Rat, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Histone H3
• Description: Rabbit Monoclonal
• Gene: H3F3A
• Conjugate: Unconjugated

Product images

All lanes : Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840) at 1/1000 dilutionLane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysateLane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Dot blot analysis of Histone H3.3 peptide (Lane 1), and Histone H3.1 peptide (Lane 2), labeled using ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840) at 1/1000 dilutionLane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysateLane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with Histone H3.3 peptideLane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with Histone H3.1 peptideLane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysateLane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with Histone H3.3 peptideLane 6 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with Histone H3.1 peptideLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

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 ab176840 (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).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3.3 with ab176840 at 1/1000 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. 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:-ve control 1: ab176840 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H3.3 with ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human colon tissue is observed. Counter stained with Hematoxylin.Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling Histone H3.3 with ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on mouse stomach tissue is observed. Counter stained with Hematoxylin.Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H3.3 with ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Rat colon tissue is observed. Counter stained with Hematoxylin.Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.