Name: Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker
Brand: RabMAb
SKU: ab109028
Rating: 5 (8 reviews)

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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: ICC, WB, IP, IHC-P, Flow Cyt
Knockout validated
Reacts with: Mouse, Rat, Human

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 & Species

Application	Species
Flow Cyt	Human
ICC	Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- ICC: HAP1 and HeLa cells; IP: MCF7 cell lysate; Flow Cyt: 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.
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
For more information see here.
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.

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

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
Flow Cyt	Human
ICC	Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

Application	Abreviews	Notes
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.
Flow Cyt		1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
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.
Flow Cyt 1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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
modifications

Phosphorylation 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.
Target information above from: UniProt accession P45973 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
Alternative names
- Antigen p25 antibody
- CBX5 antibody
- CBX5_HUMAN antibody
- CG8409 antibody
- Chromobox 5 antibody
- Chromobox homolog 5 (HP1 alpha homolog, Drosophila) antibody
- Chromobox homolog 5 antibody
- Chromobox protein homolog 5 antibody
- Epididymis luminal protein 25 antibody
- HEL25 antibody
- Heterochromatin protein 1 alpha antibody
- Heterochromatin protein 1 antibody
- Heterochromatin protein 1 homolog alpha antibody
- HP1 alpha antibody
- HP1 alpha homolog antibody
- HP1 antibody
- HP1A antibody
- HP1Hs alpha antibody
- Su(var)205 antibody
see all

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 kDa

We are unsure how to define the extra bands.
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 1/10 000 and 1/2000 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.
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)

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 - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)

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).
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)

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).
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)

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)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Publishing research using ab109028? Please let us know so that we can cite the reference in this datasheet.

ab109028 has been referenced in 5 publications.

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

Customer reviews and Q&As

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1-8 of 8 Abreviews

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (Mouse vascular smooth muscle cells)

Permeabilization

Yes - NP40

Specification

Mouse vascular smooth muscle cells

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Mar 04 2020

Application

Western blot

Sample

Mouse Cell lysate - nuclear (Mouse vascular smooth muscle cells)

Gel Running Conditions

Reduced Denaturing

Loading amount

25 µg

Specification

Mouse vascular smooth muscle cells

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Mar 04 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (U2OS)

Permeabilization

Yes - NP40

Specification

U2OS

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Mar 04 2020

Application

Western blot

Sample

Human Cell lysate - nuclear (U2OS)

Gel Running Conditions

Reduced Denaturing

Loading amount

25 µg

Specification

U2OS

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Mar 04 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (MEF)

Permeabilization

Yes - 0.5% Triton 5 minutes

Specification

MEF

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 09 2019

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (breast cancer cell lines)

Permeabilization

Yes - 0.5% Triton

Specification

breast cancer cell lines

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 01 2016

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa)

Gel Running Conditions

Reduced Denaturing

Loading amount

10 µg

Specification

HeLa

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 25 2016

Application

Western blot

Sample

Human Cell lysate - other (HeLa)

Loading amount

20 µg

Specification

HeLa

Gel Running Conditions

Reduced Denaturing

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 10 2013

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (5)

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker

Western blot abreview for Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker

Immunocytochemistry/ Immunofluorescence abreview for Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker

Western blot abreview for Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker

Immunocytochemistry/ Immunofluorescence abreview for Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker

Immunocytochemistry/ Immunofluorescence abreview for Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker

Western blot abreview for Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker

Western blot abreview for Anti-HP1 alpha antibody [EPR5777]

Post-translational
modifications