Recombinant Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)
![Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-449558-anti-hp1-alpha-antibody-epr5777-westernblot-human-mouse-rat.jpg "Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5777] to HP1 alpha - Heterochromatin marker
- Suitable for: Flow Cyt (Intra), ICC, WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker
See all HP1 alpha primary antibodies -
Description
Rabbit monoclonal [EPR5777] to HP1 alpha - Heterochromatin marker -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- ICC: HAP1 and HeLa cells; IP: MCF7 cell lysate; Flow Cyt (intra): HeLa cells; IHC-P: Human breast cancer, rat pancreas, and mouse kidney tissue sections; WB: HeLa, SH-SY5Y whole cell lysates, Rat and mouse brain lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5777 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab185018)
- HRP Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab197721)
- Alexa Fluor® 647 Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab198391)
- Alexa Fluor® 568 Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab202533)
- Alexa Fluor® 594 Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab203431)
- Alexa Fluor® 555 Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab203432)
- PE Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab210585)
- Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109028 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| ICC |
1/250.
|
|
| WB | (4) |
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
For unpurified use at 1/10000 - 1/50000. |
| IP |
1/10 - 1/100.
|
|
| IHC-P |
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
| Notes |
|---|
|
Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC
1/250. |
|
WB
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa). For unpurified use at 1/10000 - 1/50000. |
|
IP
1/10 - 1/100. |
|
IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
Target
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Function
Component of heterochromatin that recognizes and binds histone H3 tails methylated at 'Lys-9' (H3K9me), leading to epigenetic repression. In contrast, it is excluded from chromatin when 'Tyr-41' of histone H3 is phosphorylated (H3Y41ph). Can interact with lamin-B receptor (LBR). This interaction can contribute to the association of the heterochromatin with the inner nuclear membrane. Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins. -
Sequence similarities
Contains 2 chromo domains. -
Post-translational
modificationsPhosphorylation of HP1 and LBR may be responsible for some of the alterations in chromatin organization and nuclear structure which occur at various times during the cell cycle (By similarity). Phosphorylated during interphase and possibly hyper-phosphorylated during mitosis.
Ubiquitinated. -
Cellular localization
Nucleus. Chromosome. Chromosome > centromere. Component of centromeric and pericentromeric heterochromatin. Associates with chromosomes during mitosis. Associates specifically with chromatin during metaphase and anaphase. - Information by UniProt
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Database links
- Entrez Gene: 23468 Human
- Entrez Gene: 12419 Mouse
- Entrez Gene: 300266 Rat
- Omim: 604478 Human
- SwissProt: P45973 Human
- SwissProt: Q61686 Mouse
- Unigene: 349283 Human
- Unigene: 262059 Mouse
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Alternative names
- Antigen p25 antibody
- CBX5 antibody
- CBX5_HUMAN antibody
see all
Images
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Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)All lanes : Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
Lane 3 : Mouse brain lysate
Lane 4 : Rat brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 22 kDaWe are unsure how to define the extra bands.
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![Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-255523-anti-hp1-alpha-antibody-epr5777-heterochromatin-marker-western-blot.jpg)
Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: HP1 alpha knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: MCF7 cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab109028 observed at 28 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109028 was shown to specifically react with HP1 alpha when HP1 alpha knockout samples were used. Wild-type and HP1 alpha knockout samples were subjected to SDS-PAGE. ab109028 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/20 000 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 1/10000 dilution for 1 h at room temperature before imaging.
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![Immunoprecipitation - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-10-anti-hp1-alpha-antibody-epr5777-immunoprecipitation-mcf7-human.jpg)
Immunoprecipitation - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)Purified ab109028 at 1/30 dilution (2µg) immunoprecipitating HP1 alpha in MCF7 whole cell lysate.
Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab109028 + MCF7 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109028 in MCF7 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 22 kDa -
![Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-15-anti-hp1-alpha-antibody-epr5777-immunocytochemistry-hela-human.jpg)
Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HP1 alpha with Purified ab109028 at 1/250 dilution (1.22 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
![Flow Cytometry (Intracellular) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-16-anti-hp1-alpha-antibody-epr5777-flowcytometry-hela-human.jpg)
Flow Cytometry (Intracellular) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HP1 alpha with Purified ab109028 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-12-anti-hp1-alpha-antibody-epr5777-immunohistochemistry-breast-cancer-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
![Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-267408-anti-hp1-alpha-antibody-epr5777-heterochromatin-marker-immunofluorescence.jpg)
Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)ab109028 staining HP1 alpha in wild-type HAP1 cells (top panel) and HP1 alpha knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109028 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-13-anti-hp1-alpha-antibody-epr5777-immunohistochemistry-kidney-mouse.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-14-anti-hp1-alpha-antibody-epr5777-immunohistochemistry-pancreas-rat.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat pancreas tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
![Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)](https://www.abcam.com/ps/products/109/ab109028/Images/ab109028-11-benefits-of-recombinant-antibodies.png)
Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab109028 has been referenced in 8 publications.
- Lionetti MC et al. Chromatin and Cytoskeletal Tethering Determine Nuclear Morphology in Progerin-Expressing Cells. Biophys J 118:2319-2332 (2020). PubMed: 32320674
- Meseure D et al. Biopathological Significance of PIWI-piRNA Pathway Deregulation in Invasive Breast Carcinomas. Cancers (Basel) 12:N/A (2020). PubMed: 33008024
- Gomes AP et al. Dynamic Incorporation of Histone H3 Variants into Chromatin Is Essential for Acquisition of Aggressive Traits and Metastatic Colonization. Cancer Cell 36:402-417.e13 (2019). PubMed: 31564638
- Damodaran K et al. Compressive force induces reversible chromatin condensation and cell geometry dependent transcriptional response. Mol Biol Cell N/A:mbcE18040256 (2018). PubMed: 30256731
- Xie X et al. ß-Actin-dependent global chromatin organization and gene expression programs control cellular identity. FASEB J 32:1296-1314 (2018). PubMed: 29101221
- Cappuyns E et al. Mutations in ADNP affect expression and subcellular localization of the protein. Cell Cycle 17:1068-1075 (2018). PubMed: 29911927
- Pandey S et al. A novel MeCP2 acetylation site regulates interaction with ATRX and HDAC1. Genes Cancer 6:408-21 (2015). WB ; Human . PubMed: 26622943
- Robin JD et al. Telomere position effect: regulation of gene expression with progressive telomere shortening over long distances. Genes Dev 28:2464-76 (2014). ChIP . PubMed: 25403178