Specific protocols
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To our knowledge, customised protocols are not required for this product.
Please try the standard protocols listed below and let us know how you get on. |
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General protocols
Useful resources:Western blotting (WB) protocols:Immunohistochemistry (IHC) / Immunocytochemistry (ICC) protocols:Chromatin Immunoprecipitation (ChIP) protocols:Dot blot protocols:ELISA protocols:ELISPOT protocols: Flow cytometry / FACS protocols:Immunoprecipitation (IP) protocols: |
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Western blot - HP1 gamma (phospho S83) antibody (ab45270)
All lanes : Anti-HP1 gamma (phospho S83) antibody (ab45270) at 1 µg/ml
Lane 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 Lysate
Lane 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 HP1 gamma peptide (ab51840) at 1 µg/ml
Lane 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with HP1 gamma peptide (ab51840) at 1 µg/ml
Lane 10 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with HP1 gamma peptide (ab51840) at 1 µg/ml
Lane 11 : HEK293 Human embryonic kidney cell line Whole Cell Lysate with HP1 gamma peptide (ab51840) at 1 µg/ml
Lane 12 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with HP1 gamma peptide (ab51840) at 1 µg/ml
Lane 13 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate (ab28419) with HP1 gamma peptide (ab51840) at 1 µg/ml
Lane 14 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate (ab45968) with HP1 gamma peptide (ab51840) at 1 µg/ml
Lane 15 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with HP1 gamma peptide - phospho S83 (ab45386) at 1 µg/ml
Lane 16 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with HP1 gamma peptide - phospho S83 (ab45386) at 1 µg/ml
Lane 17 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with HP1 gamma peptide - phospho S83 (ab45386) at 1 µg/ml
Lane 18 : HEK293 Human embryonic kidney cell line Whole Cell Lysate with HP1 gamma peptide - phospho S83 (ab45386) at 1 µg/ml
Lane 19 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with HP1 gamma peptide - phospho S83 (ab45386) at 1 µg/ml
Lane 20 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with HP1 gamma peptide - phospho S83 (ab45386) at 1 µg/ml
Lane 21 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with HP1 gamma peptide - phospho S83 (ab45386) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 21 kDa
Observed band size : 24 kDa (why is the actual band size different from the predicted?)
Additional bands at : 40 kDa. We are unsure as to the identity of these extra bands.
Immunocytochemistry/ Immunofluorescence - HP1 gamma (phospho S83) antibody (ab45270)
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).
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