Anti-Histone H3 antibody [EPR16987] - Nuclear Loading Control and ChIP Grade

• Supplier: Abcam
• Catalog: ab176842
• Prices: $376.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EPR16987
• Applications: IHC-P ICC/IF ICC/IF WB ChIP
• Species Reactivities: Mouse, Rat, Human, Yeast, Drosophila, S. pombe, Broad species
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Histone H3 aa 100 to the C-terminus
• Description: Rabbit Monoclonal
• Gene: His3.3B
• Conjugate: Unconjugated

Product images

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Histone H3 with ab176842 at 1/2000 dilution, followed by Goat anti-rabbit Alexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/400 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 1/500 dilution and ab150120 (goat anti-mouse AlexaFluor®594 Goat secondary antibody) at 1/500 dilution (red).The negative controls are as follows;-ve control 1: ab176842 at 1/2000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. 

All lanes : Anti-Histone H3 antibody [EPR16987] - Nuclear Loading Control and ChIP Grade (ab176842) at 1 µgLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µgLane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µgLane 3 : Drosophila embryo nuclear extract (from melanogaster embryos 0-12hr) at 10 µgLane 4 : S.cerevisiae (Y190) Whole Cell Lysate at 20 µgLane 5 : S.pombe Whole Cell Lysate at 20 µgSecondaryPeroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Chromatin was prepared from HeLa 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 ab176842 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H3 with ab176842 at 1/500 dilution, followed by Goat Anti-Rabbit HRP (IgG; H&L) secondary antibody (ab97051) at 1/500 dilution. Nucleus staining on glandular epithelium of Human colon tissue is observed. Counter stained with Hematoxylin.Negative control: Used PBS instead of primary antibody, secondary antibody Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Histone H3 with ab176842 at 1/500 dilution, followed by Goat Anti-Rabbit HRP (IgG H&L) secondary antibody (ab97051) at 1/500 dilution. Nucleus staining on mouse liver tissue is observed. Counter stained with Hematoxylin.Negative control: Used PBS instead of primary antibody, secondary antibody Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling Histone H3 with ab176842 at 1/500 dilution, followed by Goat Anti-Rabbit HRP (IgG H&L) secondary antibody (ab97051) at 1/500 dilution. Nucleus staining on rat pancreas tissue is observed. Counter stained with Hematoxylin.Negative control: Used PBS instead of primary antibody, secondary antibody Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.