Anti-HP1 gamma/CBX3 (phospho S93) antibody (ab45270)
Key features and details
- Rabbit polyclonal to HP1 gamma/CBX3 (phospho S93)
- Suitable for: ICC/IF, IP, WB, IHC-P
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-HP1 gamma/CBX3 (phospho S93) antibody
See all HP1 gamma/CBX3 primary antibodies -
Description
Rabbit polyclonal to HP1 gamma/CBX3 (phospho S93) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Cow, Rhesus monkey, Orangutan -
Immunogen
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General notes
This product was previously labelled as HP1 gamma
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentrationConcentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab45270 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes ICC/IF Use a concentration of 5 µg/ml. IP Use a concentration of 5 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 21 kDa). IHC-P Use a concentration of 1 µg/ml. Target
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Function
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. -
Sequence similarities
Contains 2 chromo domains. -
Post-translational
modificationsPhosphorylated by PIM1. Phosphorylated during interphase and possibly hyper-phosphorylated during mitosis. -
Cellular localization
Nucleus. Associates with euchromatin and is largely excluded from constitutive heterochromatin. May be associated with microtubules and mitotic poles during mitosis. - Information by UniProt
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Database links
- Entrez Gene: 539788 Cow
- Entrez Gene: 11335 Human
- Entrez Gene: 12417 Mouse
- Entrez Gene: 706978 Rhesus monkey
- Omim: 604477 Human
- SwissProt: Q13185 Human
- SwissProt: P23198 Mouse
- SwissProt: Q5R6X7 Orangutan
see all -
Alternative names
- CBX 3 antibody
- CBX3 antibody
- CBX3_HUMAN antibody
see all
Images
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All lanes : Anti-HP1 gamma/CBX3 (phospho S93) 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 Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 10 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 11 : HEK293 Human embryonic kidney cell line Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 12 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 13 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate (ab28419) with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 14 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate (ab45968) with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 15 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 16 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 17 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 18 : HEK293 Human embryonic kidney cell line Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 19 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 20 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 21 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 21 kDa -
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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IHC image of HP1 gamma/CBX (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.
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HP1 gamma/CBX was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to HP1 gamma/CBX 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/CBX
Protocols
Datasheets and documents
References (6)
ab45270 has been referenced in 6 publications.
- Brenner E et al. Cancer immune control needs senescence induction by interferon-dependent cell cycle regulator pathways in tumours. Nat Commun 11:1335 (2020). PubMed: 32165639
- Poulard C et al. A post-translational modification switch controls coactivator function of histone methyltransferases G9a and GLP. EMBO Rep 18:1442-1459 (2017). PubMed: 28615290
- Lastwika KJ et al. Control of PD-L1 Expression by Oncogenic Activation of the AKT-mTOR Pathway in Non-Small Cell Lung Cancer. Cancer Res 76:227-38 (2016). PubMed: 26637667
- Rielland M et al. Senescence-associated SIN3B promotes inflammation and pancreatic cancer progression. J Clin Invest 124:2125-35 (2014). IHC-P ; Mouse . PubMed: 24691445
- Harouz H et al. Shigella flexneri targets the HP1? subcode through the phosphothreonine lyase OspF. EMBO J 33:2606-22 (2014). IHC-Fr, ICC/IF ; Guinea pig, Human . PubMed: 25216677
- Thorne JL et al. Heterochromatin protein 1 gamma and I?B kinase alpha interdependence during tumour necrosis factor gene transcription elongation in activated macrophages. Nucleic Acids Res : (2012). WB, ChIP ; Mouse . PubMed: 22649058
Images
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All lanes : Anti-HP1 gamma/CBX3 (phospho S93) 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 Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 10 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 11 : HEK293 Human embryonic kidney cell line Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 12 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 13 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate (ab28419) with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 14 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate (ab45968) with Human HP1 gamma/CBX3 peptide (ab51840) at 1 µg/ml
Lane 15 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 16 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 17 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 18 : HEK293 Human embryonic kidney cell line Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 19 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 20 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lane 21 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate withHuman HP1 gamma/CBX3 (phospho S93) peptide (ab45386) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 21 kDa -
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/CBX (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/CBX was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to HP1 gamma/CBX 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/CBX