Synthetic peptide corresponding to residues in Human HP1 gamma (Q13185).
NIH/3T3, HeLa, Molt-4 and HepG2 cell lysates; Human breast carcinoma tissue; Human normal breast tissue.
This product is a recombinant rabbit monoclonal antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated "PUR" on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/100 - 1/1000. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/10 - 1/100.
Is unsuitable for ICC.
Seems to be involved in transcriptional silencing in heterochromatin-like complexes. Recognizes and binds histone H3 tails methylated at 'Lys-9', leading to epigenetic repression. May contribute to the association of the heterochromatin with the inner nuclear membrane through its interaction with lamin B receptor (LBR). Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins.
Contains 2 chromo domains.
Phosphorylated by PIM1. Phosphorylated during interphase and possibly hyper-phosphorylated during mitosis.
Nucleus. Associates with euchromatin and is largely excluded from constitutive heterochromatin. May be associated with microtubules and mitotic poles during mitosis.
Western blot - Anti-HP1 gamma antibody [EPR10465(B)] (ab154871)
Predicted band size : 20 kDa Additional bands at : 24 kDa. We are unsure as to the identity of these extra bands.
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: HP1 gamma knockout HAP1 cell lysate (20 µg) Lane 3: NIH3T3 cell lysate (20 µg) Lane 4: HeLa cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab154871 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab154871was shown to recognize HP1 gamma when HP1 gamma knockout samples were used, along with additional cross-reactive bands. Wild-type and HP1 gamma knockout samples were subjected to SDS-PAGE. ab154871 and ab8245 (loading control to GAPDH) were both diluted to 1/500 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Overlay histogram showing HeLa cells stained with ab154871 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab154871, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Anti-HP1 gamma antibody [EPR10465(ABC)(B)] (ab154871)
All lanes : Anti-HP1 gamma antibody [EPR10465(B)] (ab154871) at 1/1000 dilution
Lane 1 : NIH/3T3 cell lysate Lane 2 : HeLa cell lysate Lane 3 : Molt-4 cell lysate Lane 4 : HepG2 cell lysate