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Rabbit Recombinant Monoclonal HP1 alpha antibody. Heterochromatin marker. Suitable for IHC-P, ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 16 publications.


Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PICC/IFIPWBFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Tested
Expected
Expected
Tested
Expected
Rat
Tested
Expected
Expected
Tested
Expected

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

For unpurified use at 1/250 - 1/500.

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Species

Rat

Dilution info

1/1000

Notes

For unpurified use at 1/250 - 1/500.

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/250 - 1/500.

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/250

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/50000.

Species

Rat

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/50000.

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/50000.

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

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.

Alternative names

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Rabbit Recombinant Monoclonal HP1 alpha antibody. Heterochromatin marker. Suitable for IHC-P, ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 16 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR5777

Purification technique

Affinity purification Protein A

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

Biological function summary

Heterochromatin protein 1 alpha acts as a modulator of gene silencing. It is part of a larger protein complex that contains various chromatin modifiers responsible for maintaining the silent state of certain genes. By mediating these interactions HP1 alpha helps regulate transcription replication timing and DNA repair processes. This regulation is essential for the structural maintenance of heterochromatin and plays a part in chromosomal stability.

Activity summary

HP1 alpha also known as HP1a or HP1 protein is a well-studied member of the heterochromatin protein 1 family. It plays a mechanical role in chromatin organization and gene regulation by binding to histone H3 when it's methylated at lysine 9 (H3K9me). This protein has a molecular mass of approximately 22-25 kDa. HP1 alpha is commonly expressed in the nuclei of eukaryotic cells where it localizes to heterochromatin regions and influences chromatin compaction.

Pathways

HP1 alpha is integral in the epigenetic regulation pathway along with other well-known proteins including SUV39H1 and SUV39H2 which are histone methyltransferases. HP1 alpha also links to the DNA damage response pathway where it recruits repair factors to maintain genomic integrity. Through these pathways HP1 alpha interacts closely with proteins like BRCA1 emphasizing its role in cellular homeostasis and response to DNA damage.

Associated diseases and disorders

Researchers associate HP1 alpha with cancer and developmental disorders. Alterations in HP1 alpha expression or function can lead to disruptions in chromatin organization contributing to oncogenesis by affecting tumor suppressor genes. Furthermore HP1 alpha links to disorders like ICF syndrome through its relationship with the DNA methylation process where it interacts with proteins such as DNMT3B. Understanding these connections provides insights into therapeutic approaches targeting epigenetic modifications in disease contexts.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunoprecipitation - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028), expandable thumbnail

    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

    All lanes: Immunoprecipitation - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (AB109028)

    Predicted band size: 22 kDa

  • Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028), expandable thumbnail

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

    We are unsure how to define the extra bands.

    All lanes: Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (AB109028) at 1/1000 dilution

    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: Mouse brain lysate at 20 µg

    Lane 4: Rat brain lysate at 20 µg

    Secondary

    All lanes: Western blot - 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.

    Purified ab109028 at 1/30 dilution (2μg) immunoprecipitating HP1 alpha in MCF7 whole cell lysate.

  • Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028), expandable thumbnail

    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.

    All lanes: Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (AB109028)

    Predicted band size: 22 kDa, 24 kDa

  • Flow Cytometry (Intracellular) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028), expandable thumbnail

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028), expandable thumbnail

    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/ Immunofluorescence - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - 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), expandable thumbnail

    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), expandable thumbnail

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028), expandable thumbnail

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

    ab109028 staining HP1 alpha in HeLa (human cervical adenocarcinoma epithelial cell). The cells were fixed with 100% methanol, permeabilized with 0.1% TritonX-100. The cells were then incubated with ab109028 at 1:100 dilution. Cells were then incubated with ab150088, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1:1000 dilution (shown in green). ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as counterstain at 1:200 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Confocal image showing nuclear staining in HeLa cell line.

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Product protocols

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