Anti-HP1 gamma (phospho S93) antibody

• Supplier: Abcam
• Catalog: ab45270
• Prices: $394.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ICC/IF ICC/IF IP WB IHC-P
• Species Reactivities: Human, Monkey, Mouse, Bovine, Orangutan
• Antigen: Synthetic peptide corresponding to Human HP1 gamma aa 50-150 conjugated to Keyhole Limpet Haemocyanin (KLH)
• Blocking Peptide: Human HP1 gamma (phospho S93) peptide
• Description: Rabbit Polyclonal
• Gene: CBX3
• Conjugate: Unconjugated

Product images

All lanes : Anti-HP1 gamma (phospho S93) antibody (ab45270) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate Lane 8 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/mlLane 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/mlLane 10 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma peptide (ab51840) at 1 µg/mlLane 11 : HEK293 Human embryonic kidney cell line Whole Cel

ICC/IF image of ab45270 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45270, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

IHC image of HP1 gamma (phospho S83) staining in Human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45270, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

HP1 gamma was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to HP1 gamma and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab45270.Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.Band: 22kDa; HP1 gamma

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